Sulfonamide compounds and uses as TNAP inhibitors

ABSTRACT

Described herein are compounds that modulate the activity of TNAP. In some embodiments, the compounds described herein inhibit TNAP. In certain embodiments, the compounds described herein are useful in the treatment of conditions associated with hyper-mineralization.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.14/379,475, filed Aug. 18, 2014, which is the National Phase entry ofInternational Application No. PCT/US2013/027191, filed Feb. 21, 2013,which claims the benefit of U.S. Provisional Application No. 61/601,957,filed Feb. 22, 2012, all of which are incorporated by reference hereinin their entireties.

STATEMENT AS TO FEDERALLY SPONSORED RESEARCH

This invention was made with government support under RC1 HL101899awarded by the National Institutes of Health. The government has certainrights in the invention.

BACKGROUND OF THE INVENTION

Among the human alkaline phosphatases, tissue-nonspecific alkalinephosphatase (TNAP) is essential for bone matrix mineralization. Thebiological function of TNAP is to hydrolyze extracellular inorganicpyrophosphate (ePP_(i)), which is an inhibitor of calcification. Lowlevels of ePP_(i) have been associated with hyper-mineralization. Thereis a need for compounds that inhibit TNAP to prevent medical conditionsassociated with hyper-mineralization, for example, osteoarthritis,medial vascular calcification, and ankylosis.

SUMMARY OF THE INVENTION

Described herein are compounds that modulate the activity of TNAP. Insome embodiments, the compounds described herein inhibit TNAP. Incertain embodiments, the compounds described herein are useful in thetreatment of conditions associated with hyper-mineralization.

In one aspect, provided herein are compounds of Formula I, or apharmaceutically acceptable salts, polymorphs, solvates, tautomers,metabolites, or N-oxides thereof:

wherein:

-   -   Y¹ and Y² are independently a bond or —N(R⁶)—, wherein at least        one of Y¹ and Y² is —N(R⁶)—;    -   L¹ and L² are independently a bond or optionally substituted        alkylene;    -   X¹ is ═N— or ═C(R²)—;    -   X² is ═N— or ═C(R³)—;    -   R¹ and R⁴ are independently selected from the group consisting        of hydrogen, —F, —Cl, —Br, —I, —CN, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹,        optionally substituted alkyl, optionally substituted cycloalkyl,        optionally substituted heterocycloalkyl, optionally substituted        alkoxy, haloalkyl, haloalkoxy, optionally substituted phenyl,        and optionally substituted 5- or 6-membered heteroaryl;    -   R², R³, and R⁵ are independently selected from the group        consisting of hydrogen, halogen, —CN, —C(O)—N(R⁷)—R⁸,        —C(O)—O—R⁹, optionally substituted alkyl, optionally substituted        cycloalkyl, optionally substituted heterocycloalkyl, optionally        substituted alkoxy, haloalkyl, haloalkoxy, optionally        substituted phenyl, and optionally substituted 5- or 6-membered        heteroaryl;    -   R⁶ is hydrogen, optionally substituted alkyl, optionally        substituted alkenyl, or optionally substituted alkynyl;    -   R⁷ and R⁸ are independently hydrogen, optionally substituted        alkyl, haloalkyl, optionally substituted cycloalkyl, optionally        substituted heterocycloalkyl, optionally substituted phenyl, or        R⁷ and R⁸ together with the nitrogen atom to which they are        attached form an optionally substituted heterocycloamino;    -   R⁹ is selected from the group consisting of hydrogen, optionally        substituted alkyl, haloalkyl, optionally substituted cycloalkyl,        optionally substituted heterocycloalkyl, and optionally        substituted phenyl; and    -   A is selected from the group consisting of —C(O)—N(R⁷)—R⁸,        —C(O)—O—R⁹, optionally substituted phenyl, and optionally        substituted 5- or 6-membered heteroaryl.

In some embodiments described above or below, provided herein arecompounds of Formula I, wherein Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; L¹ is a bond; L² is a bond; and R⁶ is hydrogen as shown inFormula Ie:

In some embodiments described above or below, provided herein arecompounds of Formula I, wherein A is optionally substituted phenyl oroptionally substituted 5- or 6-membered heteroaryl. In some embodiments,A is selected from:

wherein:

-   -   R¹² and R¹³ are independently selected from the group consisting        of hydrogen, halogen, —CN, —OH, —C(O)—N(R¹⁷)—R¹⁸, —C(O)—O—R¹⁹,        optionally substituted alkyl, optionally substituted cycloalkyl,        optionally substituted heterocycloalkyl, optionally substituted        alkoxy, haloalkyl, haloalkoxy, optionally substituted phenyl,        and optionally substituted 5- or 6-membered heteroaryl,        -   wherein:            -   R¹⁷ and R¹⁸ are independently hydrogen, optionally                substituted alkyl, haloalkyl, optionally substituted                cycloalkyl, optionally substituted heterocycloalkyl,                optionally substituted phenyl, or R¹⁷ and R¹⁸ together                with the nitrogen atom to which they are attached form                an optionally substituted heterocycloamino; and            -   R¹⁹ is selected from the group consisting of hydrogen,                optionally substituted alkyl, haloalkyl, optionally                substituted cycloalkyl, optionally substituted                heterocycloalkyl, and optionally substituted phenyl; and    -   R¹⁵ is hydrogen or optionally substituted alkyl.

In another aspect, provided herein are compounds of Formula II, orpharmaceutically acceptable salts, polymorphs, solvates, tautomers,metabolites, or N-oxides thereof:

wherein:

-   -   Y¹ and Y² are independently a bond or —N(R⁶)—, wherein at least        one of Y¹ and Y² is —N(R⁶)—;    -   L¹ and L² are independently a bond or optionally substituted        alkylene;    -   X¹ is ═N— or ═C(R²)—;    -   X² is ═N— or ═C(R³)—;    -   R¹¹ is selected from the group consisting of Cl, —CN,        —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹, optionally substituted alkyl,        optionally substituted cycloalkyl, optionally substituted        heterocycloalkyl, optionally substituted alkoxy, haloalkyl,        haloalkoxy, optionally substituted phenyl, and optionally        substituted 5- or 6-membered heteroaryl;    -   R¹⁴ is selected from the group consisting of hydrogen, Cl, Br,        —CN, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹, optionally substituted alkyl,        optionally substituted cycloalkyl, optionally substituted        heterocycloalkyl, optionally substituted alkoxy, haloalkyl,        haloalkoxy, optionally substituted phenyl, and optionally        substituted 5- or 6-membered heteroaryl;    -   R², R³, and R⁵ are independently selected from the group        consisting of hydrogen, halogen, —CN, —C(O)—N(R⁷)—R⁸,        —C(O)—O—R⁹, optionally substituted alkyl, optionally substituted        cycloalkyl, optionally substituted heterocycloalkyl, optionally        substituted alkoxy, haloalkyl, haloalkoxy, optionally        substituted phenyl, and optionally substituted 5- or 6-membered        heteroaryl;    -   R⁶ is hydrogen, optionally substituted alkyl, optionally        substituted alkenyl, or optionally substituted alkynyl;    -   R⁷ and R⁸ are independently hydrogen, optionally substituted        alkyl, haloalkyl, optionally substituted cycloalkyl, optionally        substituted heterocycloalkyl, optionally substituted phenyl, or        R⁷ and R⁸ together with the nitrogen atom to which they are        attached form an optionally substituted heterocycloamino;    -   R⁹ is selected from the group consisting of optionally        substituted alkyl, haloalkyl, optionally substituted cycloalkyl,        optionally substituted heterocycloalkyl, and optionally        substituted phenyl; and    -   A is selected from the group consisting of hydrogen, optionally        substituted alkyl, —OH, optionally substituted alkoxy,        optionally substituted haloalkoxy, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹,        optionally substituted phenyl, and optionally substituted 5- or        6-membered heteroaryl,        -   wherein:            -   if A and R⁵ are hydrogen and R¹ is methoxy, then R⁴ is                independently selected from the group consisting of                hydrogen, —Cl, —CN, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹,                optionally substituted C₂- to C₆-alkyl, optionally                substituted cycloalkyl, optionally substituted                heterocycloalkyl, optionally substituted C₂- to                C₆-alkoxy, haloalkyl, haloalkoxy, optionally substituted                phenyl, and optionally substituted 5- or 6-membered                heteroaryl.

In some embodiments described above or below, provided herein arecompounds of Formula II, wherein Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; L¹ is a bond; L² is a bond; and R⁶ is hydrogen as shown inFormula IIe:

In a further aspect, provided herein are compounds Formula III, orpharmaceutically acceptable salts, polymorphs, solvates, tautomers,metabolites, or N-oxides thereof:

wherein:

-   -   Y¹ and Y² are independently a bond or —N(R⁶)—;    -   L¹ and L² are independently a bond or optionally substituted        alkylene;    -   X¹ is ═N— or ═C(R²)—;    -   X² is ═N— or ═C(R³)—;    -   R¹¹ is selected from the group consisting of Cl, —CN,        —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹, optionally substituted alkyl,        optionally substituted cycloalkyl, optionally substituted        heterocycloalkyl, optionally substituted alkoxy, haloalkyl,        haloalkoxy, optionally substituted phenyl, and optionally        substituted 5- or 6-membered heteroaryl;    -   R¹⁴ is selected from the group consisting of hydrogen, Cl, Br,        —CN, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹, optionally substituted alkyl,        optionally substituted cycloalkyl, optionally substituted        heterocycloalkyl, optionally substituted alkoxy, haloalkyl,        haloalkoxy, optionally substituted phenyl, and optionally        substituted 5- or 6-membered heteroaryl;    -   R², R³, and R⁵ are independently selected from the group        consisting of hydrogen, halogen, —CN, —C(O)—N(R⁷)—R⁸,        —C(O)—O—R⁹, optionally substituted alkyl, optionally substituted        cycloalkyl, optionally substituted heterocycloalkyl, optionally        substituted alkoxy, haloalkyl, haloalkoxy, optionally        substituted phenyl, and optionally substituted 5- or 6-membered        heteroaryl;    -   R⁶ is hydrogen, optionally substituted alkyl, optionally        substituted alkenyl, or optionally substituted alkynyl;    -   R⁷ and R⁸ are independently hydrogen, optionally substituted        alkyl, haloalkyl, optionally substituted cycloalkyl, optionally        substituted heterocycloalkyl, optionally substituted phenyl, or        R⁷ and R⁸ together with the nitrogen atom to which they are        attached form an optionally substituted heterocycloamino;    -   R⁹ is selected from the group consisting of optionally        substituted alkyl, haloalkyl, optionally substituted cycloalkyl,        optionally substituted heterocycloalkyl, and optionally        substituted phenyl; and    -   Z is hydrogen or —N(R¹⁷)—R¹⁸, wherein:        -   if Z and R⁵ are hydrogen and R¹¹ is alkoxy, then R¹⁴ is            independently selected from the group consisting of            hydrogen, Br, —CN, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹, optionally            substituted C₂- to C₆-alkyl, optionally substituted            cycloalkyl, optionally substituted heterocycloalkyl,            optionally substituted C₂- to C₆-alkoxy, haloalkyl,            haloalkoxy, optionally substituted phenyl, and optionally            substituted 5- or 6-membered heteroaryl; and        -   R¹⁷ and R¹⁸ are independently hydrogen, optionally            substituted alkyl, haloalkyl, optionally substituted            cycloalkyl, optionally substituted heterocycloalkyl,            optionally substituted phenyl, or R¹⁷ and R¹⁸ together with            the nitrogen atom to which they are attached form an            optionally substituted heterocycloamino.

In some embodiments described above or below, provided herein arecompounds of Formula III, wherein Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; L¹ is a bond; L² is a bond; and R⁶ is hydrogen as shown inFormula IIIe:

In another aspect, provided herein are compounds Formula IV, orpharmaceutically acceptable salts, polymorphs, solvates, tautomers,metabolites, or N-oxides thereof:

wherein:

-   -   Y¹ and Y² are independently a bond or —N(R⁶)—;    -   L¹ and L² are independently a bond or optionally substituted        alkylene;    -   X¹ is ═N— or ═C(R²)—;    -   X² is ═N— or ═C(R³)—;    -   R¹ and R⁴ are independently selected from the group consisting        of hydrogen, halogen, —CN, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹,        optionally substituted alkyl, optionally substituted cycloalkyl,        optionally substituted heterocycloalkyl, optionally substituted        alkoxy, haloalkyl, haloalkoxy, optionally substituted phenyl,        and optionally substituted 5- or 6-membered heteroaryl;    -   R², R³, and R⁵ are independently selected from the group        consisting of hydrogen, halogen, —CN, —C(O)—N(R⁷)—R⁸,        —C(O)—O—R⁹, optionally substituted alkyl, optionally substituted        cycloalkyl, optionally substituted heterocycloalkyl, optionally        substituted alkoxy, haloalkyl, haloalkoxy, optionally        substituted phenyl, and optionally substituted 5- or 6-membered        heteroaryl;    -   R⁶ is hydrogen, optionally substituted alkyl, optionally        substituted alkenyl, or optionally substituted alkynyl;    -   R⁷ and R⁸ are independently hydrogen, optionally substituted        alkyl, haloalkyl, optionally substituted cycloalkyl, optionally        substituted heterocycloalkyl, optionally substituted phenyl, or        R⁷ and R⁸ together with the nitrogen atom to which they are        attached form an optionally substituted heterocycloamino;    -   R⁹ is selected from the group consisting of hydrogen, optionally        substituted alkyl, haloalkyl, optionally substituted cycloalkyl,        optionally substituted heterocycloalkyl, and optionally        substituted phenyl; and    -   W is selected from the group consisting of an optionally        substituted 5-membered heteroaryl, an optionally substituted        6-membered heteroaryl other than pyridin-3-yl, an optionally        substituted 9-membered heteroaryl, or an optionally substituted        10-membered heteroaryl other than quinolin-3-yl.

In some embodiments described above or below, provided herein arecompounds of Formula IV, wherein Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; L¹ is a bond; L² is a bond; and R⁶ is hydrogen as shown inFormula IVe:

In certain embodiments described above or below, provided herein arecompounds of Formula IV, wherein W is selected from:

wherein:

-   -   R²⁰ is selected from the group consisting of hydrogen, halogen,        —CN, —OH, —C(O)—N(R¹⁷)—R¹⁸, —C(O)—O—R¹⁹, optionally substituted        alkyl, optionally substituted cycloalkyl, optionally substituted        heterocycloalkyl, optionally substituted alkoxy, haloalkyl,        haloalkoxy, optionally substituted phenyl, and optionally        substituted 5- or 6-membered heteroaryl,        -   wherein:            -   R¹⁷ and R¹⁸ are independently hydrogen, optionally                substituted alkyl, haloalkyl, optionally substituted                cycloalkyl, optionally substituted heterocycloalkyl,                optionally substituted phenyl, or R¹⁷ and R¹⁸ together                with the nitrogen atom to which they are attached form                an optionally substituted heterocycloamino; and            -   R¹⁹ is selected from the group consisting of hydrogen,                optionally substituted alkyl, haloalkyl, optionally                substituted cycloalkyl, optionally substituted                heterocycloalkyl, and optionally substituted phenyl; and    -   R²¹ is hydrogen or optionally substituted alkyl.

In a further aspect provided herein are pharmaceutical compositionscomprising a compound of Formula I, Formula II, Formula III, or FormulaIV, or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable excipient.

In another aspect provided herein are methods of treating a disease in asubject mediated by tissue-nonspecific alkaline phosphatase (TNAP),which method comprises administering to the subject a pharmaceuticalcomposition comprising a therapeutically effective amount of a compoundof Formula I, Formula II, Formula III, or Formula IV, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable excipient. In some embodiments, the disease the disease is avascular calcification, ectopic ossification in spinal ligaments,ankylosis, or osteoarthritis. In certain embodiments, the vascularcalcification is an arterial calcification. In some embodiments, thevascular calcification is associated with diabetes mellitus I, diabetesmellitus II, idiopathic infantile arterial calcification (IIAC),Kawasaki disease, obesity, or increased age. In certain embodiments, thevascular calcification is associated with chronic renal disease (chronicrenal insufficiency), end-stage renal disease, or pre- or post-dialysisor uremia. In other embodiments, the disease is a pathologicalcalcification. In certain embodiments, the pathological calcification isankylosing spondylitis, tumoral calcinosis, fibrodysplasia ossificansprogressiva, progressive osseous heteroplasia, pseudoxanthoma elasticum,ankylosis, osteoarthritis, general arterial calcification in infancy(GACI), arterial calcification due to deficiency of CD73 (ACDC), Keutelsyndrome, peritoneal calcification, heterotopic calcification inamputees, tibial artery calcification, bone metastasis, prostheticcalcification, or Paget's disease of bone

INCORPORATION BY REFERENCE

All publications, patents, and patent applications mentioned in thisspecification are herein incorporated by reference to the same extent asif each individual publication, patent, or patent application wasspecifically and individually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF THE DRAWINGS

The novel features of the invention are set forth with particularity inthe appended claims. A better understanding of the features andadvantages of the present invention will be obtained by reference to thefollowing detailed description that sets forth illustrative embodiments,in which the principles of the invention are utilized, and theaccompanying drawings of which:

FIG. 1 is a schematic representation of the construct used to generateHprt^(ALPL) transgenic mice. The human ALPL cDNA is driven by theubiquitous CAG promoter only when Cre-recombinase has excised theloxP-flanked STOP cassette. In recombined ES cells, Hprt isreconstituted by introduction of the human promoter and exons. The humangrowth hormone polyA signal was fused to the end of the ALPL cDNA.Diagram is not depicted to scale.

FIG. 2 illustrates the characterization of the phenotype of[Hprt^(ALPL/Y); Tagln-Cre^(+/−)] mice. Top panels show histochemicalstaining for alkaline phosphatase activity (ALP), calcium deposition(Alizarin Red) and phosphate deposition (von Kossa). The lower panelsshow imaging of aortic calcification by X-ray and micro-computedtomography (μCT) and by whole-mount Alizarin Red staining.

FIG. 3 illustrates the survival curve and heart size in male[Hprt^(ALPL/Y); Tagln-Cre^(+/−)] mice. A) Survival curve is based on 17male mice. B) Heart size at autopsy at 37 days of age.

DETAILED DESCRIPTION OF THE INVENTION

Alkaline phosphatases (APs) are dimeric enzymes present in mostorganisms (Millan J L 2006, Wiley-VCH Verlag GmbH & Co, Weinheim,Germany pp. 1-322). They catalyze the hydrolysis of phosphomonoesterswith release of inorganic phosphate (Pi) and alcohol. In humans, threeof the four isozymes are tissue-specific, i.e., the intestinal (IAP),placental (PLAP), and germ cell (GCAP) APs.

The fourth AP is tissue-nonspecific (TNAP or ALPL) and is expressed inbone, liver and kidney. Specifically, TNAP is expressed on the cellmembranes of hypertrophic chrondrocytes, osteoblasts, and odontoblastsand is concentrated on the membranes of matrix vesicles budding fromthese cells. (Hoshi K, Amizuka N, Oda K, Ikehara Y, Ozawa H, HistochemCell Biol 1997: 107:183-191; Miao D, Scutt A, H Histochem Cytochem 2002;50: 333-340). TNAP has been found to hydrolyze extracellularpyrophosphate (ePPi) during the process of bone mineralization. (JohnsonK A, Hessle L, Wennberg C, Mauro S, Narisawa S, Goding J, Sano K, MillanJ L, Terkeltaub R 2000; Am J Phys Regulatory and Integrative Physiology279: 81365-1377-17; Hessle L, Johnson K A, Anderson H C, Narisawa S,Sali A, Goding J W, Terkeltaub R, Millan J L 2002; Proc Natl Acad SciUSA 99:9445-9449; Johnson K, Goding J, Van Etten D, Sali A, Hu S I,Farley D, Krug H, Hessle L, Millan J L, Terkeltaub R 2003; J Bone MinRes 18:994-1004). This decreases the amount of ePPi, which is aninhibitor of hydroxyapatite formation, and provides phosphate (Pi) forthe formation of hydroxyapatite. Thus, TNAP is an important player inbone generation, as the balance between ePPi and Pi is critical inmineralization. (Terkeltaub R A, Am J Physiol Cell Physiol 2001; 281:C1-C11).

Physiological calcification occurs in hard tissues, i.e., bone,growth-plate cartilage and dentin as part of normal development andmaintenance of the skeletal system. Pathological calcification occurs insoft tissues, such as articular cartilage, cardiovascular tissues, thekidney, skin, muscles and tendon. (Kirsch T, Curr Opin Rhematol 2006:18: 174-180). “Pathological calcification,” as used herein, refers toany formation, growth or deposition of extracellular matrixhydroxyapatite (calcium phosphate) crystal deposits in any tissue otherthan bone, growth-plate cartilage and dentin, or to calcification inbone, growth-plate cartilage and dentin that is not part of normaldevelopment and maintenance of the skeletal system. Examples ofdisorders involving pathological calcification include ankylosingspondylitis, tumoral calcinosis, fibrodysplasia ossificans progressiva,progressive osseous heteroplasia, and pseudoxanthoma elasticum. Otherconditions involving pathological calcification include ankylosis,osteoarthritis, general arterial calcification in infancy (GACI),arterial calcification due to deficiency of CD73 (ACDC), and Keutelsyndrome. Pathological calcification has also been found to occur inperitoneal calcification, heterotopic calcification in amputees, tibialartery calcification, bone metastasis, prosthetic calcification, andPaget's disease of bone.

Vascular calcification is the most common form of pathologicalcalcification. “Vascular calcification,” as used herein, refers toformation, growth or deposition of extracellular matrix hydroxyapatite(calcium phosphate) crystal deposits in blood vessels. Vascularcalcification encompasses coronary, valvular, aortic, and other bloodvessel calcification. The term includes atherosclerotic and medial wallcalcification.

TNAP has been found to play a role in pathological calcification ofvascular tissues. Increased expression of TNAP has been found toaccelerate calcification by bovine vascular smooth muscle cells (VSMCs)(Shioi A, Nishizawa Y, Jono S, Koyama H, Hosoi M, Morii H 1995,Arterioscler Thromb Vase Biol 15:2003-2009), and TNAP-rich vesicles arefound at sites of mineralization in human arteries (Hsu H H, Camacho N P1999, Atherosclerosis 143:353-362; Hui M, Li S Q, Holmyard D, Cheng P1997, Calcified Tissue International 60:467-72.; Hui M, Tenenbaum H C1998, Anatomical Record 253:91-94. Tanimura A, McGregor D H, Anderson HC 1986, J Exp Pathol 2:261-273. Tanimura A, McGregor D H, Anderson H C1986, J Exp Pathol 2:275-297). In addition, calcification of rat aortaand human valve interstitial cells in culture has been shown to bedependent on TNAP activity (Lomashvili K, Cobbs S, Hennigar R,Hardcastle K, O'Neill W C 2004, J Am. Soc. Nephrol. 15: 1392-1401;Mathieu P, Voisine P, Pepin A, Shetty R, Savard N, Dagenais F 2005, JHeart Valve Disease 14:353-357).

Vascular calcification is a well-recognized and common complication ofchronic kidney disease (CKD) (Giachelli, C. J. Am. Soc. Nephrol. 15:2959-64, 2004; Raggi, P. et al. J. Am. Coll. Cardiol. 39: 695-701,2002). Studies show that abnormalities in calcium and phosphorusmetabolism, resulting in increased HA deposition contribute to thedevelopment of arterial calcification, and to cardiovascular disease, inpatients with end-stage renal disease (Goodman, W. et al. N. Engl. J.Med. 342: 1478-83, 2000; Guerin, A. et al. Nephrol. Dial. Transplant15:1014-21, 2000; Vattikuti, R. & Towler, D. Am. J. Physiol. Endocrinol.Metab. 286: E686-96, 2004). While the causes of vascular calcificationin CKD remain to be elucidated, associated risk factors include age,gender, hypertension, time on dialysis, diabetes and glucoseintolerance, obesity, and cigarette smoking (Zoccali C. Nephrol. Dial.Transplant 15: 454-7, 2000). These conventional risk factors, however,do not adequately explain the high mortality rates from cardiovascularcauses in the patient population.

CKD is generally accompanied by secondary hyperparathyroidism (HPT). HPTis characterized by elevated parathyroid hormone (PTH) serum levels anddisordered mineral metabolism. Elevations in serum calcium, phosphorus,and HA in patients with secondary HPT have been associated with anincreased risk of vascular calcification (Chertow, G. et al. Kidney Int.62: 245-52, 2002; Goodman, W. et al. N. Engl. J. Med. 342: 1478-83,2000; Raggi, P. et al. J. Am. Coll. Cardiol. 39: 695-701, 2002).Commonly used therapeutic interventions for secondary HPT, such ascalcium-based phosphate binders and doses of active vitamin D sterolscan result in hypercalcemia and hyperphosphatemia (Chertow, G. et al.Kidney hit. 62: 245-52, 2002; Tan, A. et al. Kidney Int 51: 317-23,1997; Gallieni, M. et al. Kidney Int 42: 1191-8, 1992), which areassociated with the development or exacerbation of vascularcalcification.

Some patients with end-stage renal disease develop a severe form ofocclusive arterial disease called calciphylaxis or calcific uremicarteriolopathy. This syndrome is characterized by extensive calciumdeposition in small arteries (Gipstein R. et al. Arch Intern Med 136:1273-80, 1976; Richens G. et al. J Am Acad. Dermatol. 6: 537-9, 1982).In patients with this disease, arterial calcification and vascularocclusion lead to tissue ischemia and necrosis. Involvement ofperipheral vessels can cause ulceration of the skin of the lower legs organgrene of the digits of the feet or hands. Ischemia and necrosis ofthe skin and subcutaneous adipose tissue of the abdominal wall, thighsand/or buttocks are features of a proximal form of calcific uremicarteriolopathy (Budisavljevic M. et al. J Am Soc Nephrol. 7: 978-82,1996; Ruggian J. et al. Am. J. Kidney Dis. 28: 409-14, 1996).

“Atherosclerotic calcification” refers vascular calcification occurringin atheromatous plaques along the intimal layer of arteries.Atherosclerotic calcification is associated with lipid-laden macrophagesand intimal hyperplasia. “Medial calcification,” “medial wallcalcification,” or “Monckeberg's sclerosis,” as used herein, meanscalcification characterized by the presence of calcium in the medialwall of arteries. Medial calcification occurs in the media of a bloodvessel in conjunction with a phenotypic transformation of smooth musclecells into osteoblast-like cells.

Both forms of vascular calcification are associated with variousdiseases and disorders. For instance, both atherosclerotic and medialcalcification has been found to be common in uremic patients (Proudfoot,D & Shanahan, C. Herz 26: 245-51, 2001; Chen, N. & Moe, S. Semin Nephrol24: 61-8, 2004) and in patients with diabetes mellitus I and II.Conditions characterized by medial wall calcification include idiopathicinfantile arterial calcification (IIAC), Kawasaki disease, end-stagerenal disease, diabetes, and obesity. Medial wall calcification is alsoa general characteristic of increased age.

Atherosclerotic calcification is usually greatest in large,well-developed lesions, and such lesions have been found to increasewith age (Wexler L. et al. Circulation 94: 1175-92, 1996; Rumberger J.et al. Mayo Clin Proc 1999; 74: 243-52.). The extent of atheroscleroticcalcification in patients with atherosclerosis generally corresponds toseverity of disease. Unlike medial wall calcification, atheroscleroticvascular lesions, whether or not they contain calcium, impinge upon thearterial lumen and compromise blood flow. The localized deposition ofcalcium within atherosclerotic plaques likely occurs because ofinflammation due to oxidized lipids and other oxidative stresses andinfiltration by monocytes and macrophages (Berliner J. et al.Circulation 91: 2488-96, 1995).

Current therapies to normalize serum mineral levels or to decrease,inhibit, or prevent extraskeletal calcification are of limited efficacyand cause unacceptable side effects. Therefore, there exists a need foran effective method of inhibiting and preventing extraskeletalcalcification.

Due to its role in hydrolyzing ePPi, inhibiting TNAP function reducespathological calcification. In some embodiments, administration of acompound of Formula I-IV retards, reverses, or prevents the formation,growth or deposition of extracellular matrix hydroxyapatite crystaldeposits. In certain embodiments, the invention provides a method ofinhibiting, decreasing or preventing pathological calcification in anindividual. In some embodiments, the invention provides a method ofinhibiting, decreasing or preventing vascular calcification in anindividual. In some embodiments, the present invention provides a methodof treating or preventing atherosclerotic calcification, medialcalcification, vascular calcification associated with diabetes mellitusI and II, idiopathic infantile arterial calcification (IIAC), Kawasakidisease, obesity, and/or increased age. In some embodiments, theinvention provides a method of inhibiting, decreasing or preventingvascular calcification associated with chronic renal disease (chronicrenal insufficiency) or end-stage renal disease.

Definitions

In the following description, certain specific details are set forth inorder to provide a thorough understanding of various embodiments.However, one skilled in the art will understand that the invention maybe practiced without these details. In other instances, well-knownstructures have not been shown or described in detail to avoidunnecessarily obscuring descriptions of the embodiments. Unless thecontext requires otherwise, throughout the specification and claimswhich follow, the word “comprise” and variations thereof, such as,“comprises” and “comprising” are to be construed in an open, inclusivesense, that is, as “including, but not limited to.” Further, headingsprovided herein are for convenience only and do not interpret the scopeor meaning of the claimed invention.

Reference throughout this specification to “one embodiment” or “anembodiment” means that a particular feature, structure or characteristicdescribed in connection with the embodiment is included in at least oneembodiment. Thus, the appearances of the phrases “in one embodiment” or“in an embodiment” in various places throughout this specification arenot necessarily all referring to the same embodiment. Furthermore, theparticular features, structures, or characteristics may be combined inany suitable manner in one or more embodiments. Also, as used in thisspecification and the appended claims, the singular forms “a,” “an,” and“the” include plural referents unless the content clearly dictatesotherwise. It should also be noted that the term “or” is generallyemployed in its sense including “and/or” unless the content clearlydictates otherwise.

The terms below, as used herein, have the following meanings, unlessindicated otherwise:

“Amino” refers to the —NH₂ radical.

“Cyano” or “nitrile” refers to the —CN radical.

“Hydroxy” or “hydroxyl” refers to the —OH radical.

“Nitro” refers to the —NO₂ radical.

“Oxo” refers to the ═O substituent.

“Thioxo” refers to the ═S substituent.

“Alkyl” refers to a straight or branched hydrocarbon chain radical,which is fully saturated or comprises unsaturations, has from one tothirty carbon atoms, and is attached to the rest of the molecule by asingle bond. Alkyls comprising any number of carbon atoms from 1 to 30are included. An alkyl comprising up to 30 carbon atoms is refereed toas a C₁-C₃₀ alkyl, likewise, for example, an alkyl comprising up to 12carbon atoms is a C₁-C₁₂ alkyl. Alkyls (and other moieties definedherein) comprising other numbers of carbon atoms are representedsimilarly. Alkyl groups include, but are not limited to, C₁-C₃₀ alkyl,C₁-C₂₀ alkyl, C₁-C₁₅ alkyl, C₁-C₁₀ alkyl, C₁-C₈ alkyl, C₁-C₆ alkyl,C₁-C₄ alkyl, C₁-C₃ alkyl, C₁-C₂ alkyl, C₂-C₈ alkyl, C₃-C₈ alkyl andC₄-C₈ alkyl. Representative alkyl groups include, but are not limitedto, methyl, ethyl, n-propyl, 1-methylethyl (iso-propyl), n-butyl,i-butyl, s-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl,2-methylhexyl, vinyl, allyl, propynyl, and the like. Alkyl comprisingunsaturations include alkenyl and alkynyl groups. Unless statedotherwise specifically in the specification, an alkyl group may beoptionally substituted as described below.

“Alkylene” or “alkylene chain” refers to a straight or branched divalenthydrocarbon chain, as described for alkyl above. Unless stated otherwisespecifically in the specification, an alkylene group may be optionallysubstituted as described below.

“Alkoxy” refers to a radical of the formula —OR_(a) where R_(a) is analkyl radical as defined. Unless stated otherwise specifically in thespecification, an alkoxy group may be optionally substituted asdescribed below.

“Aryl” refers to a radical derived from a hydrocarbon ring systemcomprising hydrogen, 6 to 30 carbon atoms and at least one aromaticring. The aryl radical may be a monocyclic, bicyclic, tricyclic ortetracyclic ring system, which may include fused or bridged ringsystems. Aryl radicals include, but are not limited to, aryl radicalsderived from the hydrocarbon ring systems of aceanthrylene,acenaphthylene, acephenanthrylene, anthracene, azulene, benzene,chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane,indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene, andtriphenylene. Unless stated otherwise specifically in the specification,the term “aryl” or the prefix “ar-” (such as in “aralkyl”) is meant toinclude aryl radicals that are optionally substituted.

“Cycloalkyl” refers to a stable, non-aromatic, monocyclic or polycycliccarbocyclic ring, which may include fused or bridged ring systems, whichis saturated or unsaturated, and attached to the rest of the molecule bya single bond. Representative cycloalkyls include, but are not limitedto, cycloaklyls having from three to fifteen carbon atoms, from three toten carbon atoms, from three to eight carbon atoms, from three to sixcarbon atoms, from three to five carbon atoms, or three to four carbonatoms. Monocyclic cyclcoalkyl radicals include, for example,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, andcyclooctyl. Polycyclic radicals include, for example, adamantyl,norbornyl, decalinyl, and 7,7-dimethyl-bicyclo[2.2.1]heptanyl. Unlessotherwise stated specifically in the specification, a cycloalkyl groupmay be optionally substituted. Illustrative examples of cycloalkylgroups include, but are not limited to, the following moieties:

and the like.

“Fused” refers to any ring structure described herein which is fused toan existing ring structure. When the fused ring is a heterocyclyl ringor a heteroaryl ring, any carbon atom on the existing ring structurewhich becomes part of the fused heterocyclyl ring or the fusedheteroaryl ring may be replaced with a nitrogen atom.

“Halo” or “halogen” refers to bromo, chloro, fluoro or iodo.

“Haloalkyl” refers to an alkyl radical, as defined above, that issubstituted by one or more halo radicals, as defined above, e.g.,trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl,2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl,1,2-dibromoethyl, and the like. Unless stated otherwise specifically inthe specification, a haloalkyl group may be optionally substituted.

“Haloalkoxy” similarly refers to a radical of the formula —OR_(a) whereR_(a) is a haloalkyl radical as defined. Unless stated otherwisespecifically in the specification, a haloalkoxy group may be optionallysubstituted as described below.

“Heteroycycloalkyl” or “heterocyclyl” or “heterocyclic ring” refers to astable 3- to 24-membered non-aromatic ring radical comprising 2 to 23carbon atoms and from one to 8 heteroatoms selected from the groupconsisting of nitrogen, oxygen, phosphorous and sulfur. Unless statedotherwise specifically in the specification, the heterocyclyl radicalmay be a monocyclic, bicyclic, tricyclic or tetracyclic ring system,which may include fused or bridged ring systems; and the nitrogen,carbon or sulfur atoms in the heterocyclyl radical may be optionallyoxidized; the nitrogen atom may be optionally quaternized; and theheterocyclyl radical may be partially or fully saturated. Examples ofsuch heterocyclyl radicals include, but are not limited to, dioxolanyl,thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl,imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl,octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl,piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl,thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl,thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl,1,1-dioxo-thiomorpholinyl, 12-crown-4, 15-crown-5, 18-crown-6,21-crown-7, aza-18-crown-6, diaza-18-crown-6, aza-21-crown-7, anddiaza-21-crown-7. Unless stated otherwise specifically in thespecification, a heterocyclyl group may be optionally substituted.Illustrative examples of heterocycloalkyl groups, also referred to asnon-aromatic heterocycles, include:

and the like. The term heterocycloalkyl also includes all ring forms ofthe carbohydrates, including but not limited to the monosaccharides, thedisaccharides and the oligosaccharides. Unless otherwise noted,heterocycloalkyls have from 2 to 10 carbons in the ring. It isunderstood that when referring to the number of carbon atoms in aheterocycloalkyl, the number of carbon atoms in the heterocycloalkyl isnot the same as the total number of atoms (including the heteroatoms)that make up the heterocycloalkyl (i.e. skeletal atoms of theheterocycloalkyl ring). Unless stated otherwise specifically in thespecification, a heterocycloalkyl group may be optionally substituted.

“Heteroaryl” refers to a 5- to 14-membered ring system radicalcomprising hydrogen atoms, one to thirteen carbon atoms, one to sixheteroatoms selected from the group consisting of nitrogen, oxygen,phosphorous and sulfur, and at least one aromatic ring. For purposes ofthis invention, the heteroaryl radical may be a monocyclic, bicyclic,tricyclic or tetracyclic ring system, which may include fused or bridgedring systems; and the nitrogen, carbon or sulfur atoms in the heteroarylradical may be optionally oxidized; the nitrogen atom may be optionallyquaternized. Examples include, but are not limited to, azepinyl,acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl,benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl,benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl,benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl,benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl(benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl,carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl,furanonyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl,isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl,isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl,oxiranyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl,1-oxidopyridazinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl,phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl,pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl,quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl,tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl,triazinyl, and thiophenyl (i.e., thienyl). Unless stated otherwisespecifically in the specification, a heteroaryl group may be optionallysubstituted.

All the above groups may be either substituted or unsubstituted. Theterm “substituted” as used herein means any of the above groups (e.g,alkyl, alkylene, alkoxy, aryl, cycloalkyl, haloalkyl, heterocyclyland/or heteroaryl) may be further functionalized wherein at least onehydrogen atom is replaced by a bond to a non-hydrogen atom substituent.Unless stated specifically in the specification, a substituted group mayinclude one or more substituents selected from: oxo, —CO₂H, nitrile,nitro, hydroxyl, thiooxy, alkyl, alkylene, alkoxy, aryl, cycloalkyl,heterocyclyl, heteroaryl, dialkylamines, arylamines, alkylarylamines,diarylamines, trialkylammonium (—N⁺R₃), N-oxides, imides, and enamines;a silicon atom in groups such as trialkylsilyl groups, dialkylarylsilylgroups, alkyldiarylsilyl groups, triarylsilyl groups, perfluoroalkyl orperfluoroalkoxy, for example, trifluoromethyl or trifluoromethoxy.“Substituted” also means any of the above groups in which one or morehydrogen atoms are replaced by a higher-order bond (e.g., a double- ortriple-bond) to a heteroatom such as oxygen in oxo, carbonyl, carboxyl,and ester groups; and nitrogen in groups such as imines, oximes,hydrazones, and nitriles. For example, “substituted” includes any of theabove groups in which one or more hydrogen atoms are replaced with—NR_(g)C(═O)NR_(g)R_(h), —NR_(g)C(═O)OR_(h), —NR_(g)SO₂R_(h),—OC(═O)NR_(g)R_(h), —OR_(g), —SR_(g), —SOR_(g), —SO₂R_(g), —OSO₂R_(g),—SO₂OR_(g), ═NSO₂R_(g), and —SO₂NR_(g)R_(h). “Substituted” also meansany of the above groups in which one or more hydrogen atoms are replacedwith —C(═O)R_(g), —C(═O)OR_(g), —CH₂SO₂R_(g), —CH₂SO₂NR_(g)R_(h), —SH,—SR_(g) or —SSR_(g). In the foregoing, R_(g) and R_(h) are the same ordifferent and independently hydrogen, alkyl, alkoxy, alkylamino,thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl,heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl,N-heteroaryl and/or heteroarylalkyl. In addition, each of the foregoingsubstituents may also be optionally substituted with one or more of theabove substituents. Furthermore, any of the above groups may besubstituted to include one or more internal oxygen, sulfur, or nitrogenatoms. For example, an alkyl group may be substituted with one or moreinternal oxygen atoms to form an ether or polyether group. Similarly, analkyl group may be substituted with one or more internal sulfur atoms toform a thioether, disulfide, etc.

The term “optional” or “optionally” means that the subsequentlydescribed event or circumstance may or may not occur, and that thedescription includes instances where said event or circumstance occursand instances in which it does not. For example, “optionally substitutedalkyl” means either “alkyl” or “substituted alkyl” as defined above.Further, an optionally substituted group may be un-substituted (e.g.,—CH₂CH₃), fully substituted (e.g., —CF₂CF₃), mono-substituted (e.g.,—CH₂CH₂F) or substituted at a level anywhere in-between fullysubstituted and mono-substituted (e.g., —CH₂CHF₂, —CH₂CF₃, —CF₂CH₃,—CFHCHF₂, etc). It will be understood by those skilled in the art withrespect to any group containing one or more substituents that suchgroups are not intended to introduce any substitution or substitutionpatterns (e.g., substituted alkyl includes optionally substitutedcycloalkyl groups, which in turn are defined as including optionallysubstituted alkyl groups, potentially ad infinitum) that are stericallyimpractical and/or synthetically non-feasible. Thus, any substituentsdescribed should generally be understood as having a maximum molecularweight of about 1,000 daltons, and more typically, up to about 500daltons.

An “effective amount” or “therapeutically effective amount” refers to anamount of a compound administered to a mammalian subject, either as asingle dose or as part of a series of doses, which is effective toproduce a desired therapeutic effect.

“Treatment” of an individual (e.g. a mammal, such as a human) or a cellis any type of intervention used in an attempt to alter the naturalcourse of the individual or cell. In some embodiments, treatmentincludes administration of a pharmaceutical composition, subsequent tothe initiation of a pathologic event or contact with an etiologic agentand includes stabilization of the condition (e.g., condition does notworsen, e.g., cancer does not metastasize and the like) or alleviationof the condition (e.g., reduction in tumor size, remission of cancer,absence of symptoms of autoimmune disease and the like). In otherembodiments, treatment also includes prophylactic treatment (e.g.,administration of a composition described herein when an individual issuspected to be suffering from a condition described herein).

A “tautomer” refers to a proton shift from one atom of a molecule toanother atom of the same molecule. The compounds presented herein mayexist as tautomers. Tautomers are compounds that are interconvertible bymigration of a hydrogen atom, accompanied by a switch of a single bondand adjacent double bond. In bonding arrangements where tautomerizationis possible, a chemical equilibrium of the tautomers will exist. Alltautomeric forms of the compounds disclosed herein are contemplated. Theexact ratio of the tautomers depends on several factors, includingtemperature, solvent, and pH. Some examples of tautomericinterconversions include:

A “metabolite” of a compound disclosed herein is a derivative of thatcompound that is formed when the compound is metabolized. The term“active metabolite” refers to a biologically active derivative of acompound that is formed when the compound is metabolized. The term“metabolized,” as used herein, refers to the sum of the processes(including, but not limited to, hydrolysis reactions and reactionscatalyzed by enzymes, such as, oxidation reactions) by which aparticular substance is changed by an organism. Thus, enzymes mayproduce specific structural alterations to a compound. For example,cytochrome P450 catalyzes a variety of oxidative and reductive reactionswhile uridine diphosphate glucuronyl transferases catalyze the transferof an activated glucuronic-acid molecule to aromatic alcohols, aliphaticalcohols, carboxylic acids, amines and free sulfhydryl groups. Furtherinformation on metabolism may be obtained from The Pharmacological Basisof Therapeutics, 9th Edition, McGraw-Hill (1996). Metabolites of thecompounds disclosed herein can be identified either by administration ofcompounds to a host and analysis of tissue samples from the host, or byincubation of compounds with hepatic cells in vitro and analysis of theresulting compounds. Both methods are well known in the art. In someembodiments, metabolites of a compound are formed by oxidative processesand correspond to the corresponding hydroxy-containing compound. In someembodiments, a compound is metabolized to pharmacologically activemetabolites.

Compounds

Described herein are compounds that modulate the activity of TNAP. Insome embodiments, the compounds described herein inhibit TNAP. Incertain embodiments, the compounds described herein are useful in thetreatment of conditions associated with hyper-mineralization.

In one aspect, provided herein are compounds of Formula I, or apharmaceutically acceptable salts, polymorphs, solvates, tautomers,metabolites, or N-oxides thereof:

wherein:

-   -   Y¹ and Y² are independently a bond or —N(R⁶)—, wherein at least        one of Y¹ and Y² is —N(R⁶)—;    -   L¹ and L² are independently a bond or optionally substituted        alkylene;    -   X¹ is ═N— or ═C(R²)—;    -   X² is ═N— or ═C(R³)—;    -   R¹ and R⁴ are independently selected from the group consisting        of hydrogen, halogen, —CN, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹,        optionally substituted alkyl, optionally substituted cycloalkyl,        optionally substituted heterocycloalkyl, optionally substituted        alkoxy, haloalkyl, haloalkoxy, optionally substituted phenyl,        and optionally substituted 5- or 6-membered heteroaryl;    -   R², R³, and R⁵ are independently selected from the group        consisting of hydrogen, halogen, —CN, —C(O)—N(R⁷)—R⁸,        —C(O)—O—R⁹, optionally substituted alkyl, optionally substituted        cycloalkyl, optionally substituted heterocycloalkyl, optionally        substituted alkoxy, haloalkyl, haloalkoxy, optionally        substituted phenyl, and optionally substituted 5- or 6-membered        heteroaryl;    -   R⁶ is hydrogen, optionally substituted alkyl, optionally        substituted alkenyl, or optionally substituted alkynyl;    -   R⁷ and R⁸ are independently hydrogen, optionally substituted        alkyl, haloalkyl, optionally substituted cycloalkyl, optionally        substituted heterocycloalkyl, optionally substituted phenyl, or        R⁷ and R⁸ together with the nitrogen atom to which they are        attached form an optionally substituted heterocycloamino;    -   R⁹ is selected from the group consisting of hydrogen, optionally        substituted alkyl, haloalkyl, optionally substituted cycloalkyl,        optionally substituted heterocycloalkyl, and optionally        substituted phenyl; and    -   A is selected from the group consisting of —C(O)—N(R⁷)—R⁸,        —C(O)—O—R⁹, optionally substituted phenyl, and optionally        substituted 5- or 6-membered heteroaryl.

In some embodiments described above or below, provided herein arecompounds of Formula I, wherein Y¹ is a bond and Y² is —N(R⁶)— as shownin Formula (Ia):

In certain embodiments described above or below, provided herein arecompounds of Formula I, wherein Y¹ is a bond and Y² is —N(R⁶)—; and X²is ═C(R³)— as shown in Formula (Ib):

In some embodiments described above or below, provided herein arecompounds of Formula I, wherein Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; and L¹ is a bond as shown in Formula Ic:

In certain embodiments described above or below, provided herein arecompounds of Formula I, wherein Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; L¹ is a bond; and L² is a bond as shown in Formula Id:

In some embodiments described above or below, provided herein arecompounds of Formula I, wherein Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; L¹ is a bond; L² is a bond; and R⁶ is hydrogen as shown inFormula Ie:

For any embodiments described above or below, provided herein arecompounds of Formula I, wherein L² is an optionally substitutedalkylene.

For any embodiments described above or below, provided herein arecompounds of Formula I, wherein X¹ is ═C(R²)—.

For any embodiments described above or below, provided herein arecompounds of Formula I, wherein R², R³, and R⁵ are independentlyselected from the group consisting of hydrogen, —F, —Cl, —Br, —CN,—C(O)—OMe, methyl, —OMe, and —OCF₃. In some embodiments, R², R³, and R⁵are independently selected from the group consisting of hydrogen, Cl,methyl, and —OMe. In certain embodiments, R² and R³ are hydrogen.

For any embodiments described above or below, provided herein arecompounds of Formula I, wherein R¹ and R⁴ are independently selectedfrom the group consisting of hydrogen, —F, —Cl, —Br, —CN,—C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹, methyl, —OMe, —OCF₃, optionally substitutedphenyl, and optionally substituted 5- or 6-membered heteroaryl. In someembodiments, R⁴ is optionally substituted phenyl or optionallysubstituted 5- or 6-membered heteroaryl. In certain embodiments, R⁴ is—C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹. In some embodiments, R¹ and R⁴ areindependently selected from the group consisting of —F, —Cl, —Br, —CN,—OMe, and —OCF₃. In certain embodiments, R¹ is —OMe or —OCF₃. In someembodiments, R¹ is —OMe and R⁴ is —Cl.

In some embodiments described above or below, provided herein arecompounds of Formula I, wherein A is optionally substituted phenyl oroptionally substituted 5- or 6-membered heteroaryl. In some embodiments,A is selected from:

wherein:

-   -   R¹² and R¹³ are independently selected from the group consisting        of hydrogen, halogen, —CN, —OH, —C(O)—N(R¹⁷)—R¹⁸, —C(O)—O—R¹⁹,        optionally substituted alkyl, optionally substituted cycloalkyl,        optionally substituted heterocycloalkyl, optionally substituted        alkoxy, haloalkyl, haloalkoxy, optionally substituted phenyl,        and optionally substituted 5- or 6-membered heteroaryl,        -   wherein:            -   R¹⁷ and R¹⁸ are independently hydrogen, optionally                substituted alkyl, haloalkyl, optionally substituted                cycloalkyl, optionally substituted heterocycloalkyl,                optionally substituted phenyl, or R¹⁷ and R¹⁸ together                with the nitrogen atom to which they are attached form                an optionally substituted heterocycloamino; and            -   R¹⁹ is selected from the group consisting of hydrogen,                optionally substituted alkyl, haloalkyl, optionally                substituted cycloalkyl, optionally substituted                heterocycloalkyl, and optionally substituted phenyl; and    -   R¹⁵ is hydrogen or optionally substituted alkyl.

In certain embodiments described above or below, provided herein arecompounds of Formula I, wherein A is

wherein R¹² and R¹³ are independently selected from the group consistingof hydrogen, —F, —CN, —OH, —OMe, and —C(O)—O-Me.

In certain embodiments described above or below, provided herein arecompounds of Formula I, wherein A is

wherein R¹² and R¹³ are independently selected from the group consistingof hydrogen, —F, —CN, —OH, —OMe, and —C(O)—O-Me.

In certain embodiments described above or below, provided herein arecompounds of Formula I, wherein A is

wherein R¹² and R¹³ are independently selected from the group consistingof hydrogen, —F, —CN, —OH, —OMe, and —C(O)—O-Me.

In certain embodiments described above or below, provided herein arecompounds of Formula I, wherein A is

wherein R¹² and R¹³ are independently selected from the group consistingof hydrogen, —F, —CN, —OH, —OMe, and —C(O)—O-Me.

In certain embodiments described above or below, provided herein arecompounds of Formula I, wherein A is

wherein R¹² and R¹³ are independently selected from the group consistingof hydrogen, —F, —CN, —OH, —OMe, and —C(O)—O-Me.

In some embodiments described above or below, provided herein arecompounds of Formula I, wherein A is —C(O)—O—R⁹. In certain embodiments,R⁹ is selected from hydrogen, optionally substituted alkyl, optionallysubstituted cycloalkyl, and optionally substituted phenyl. In someembodiments, R⁹ is selected from hydrogen, methyl, ethyl, propyl,cyclohexyl, and phenyl.

In certain embodiments described above or below, provided herein arecompounds of Formula I, wherein A is —C(O)—N(R⁷)—R⁸. In someembodiments, R⁷ and R⁸ together with the nitrogen atom to which they areattached form an optionally substituted heterocycloamino. In certainembodiments, the optionally substituted heterocycloamino is anoptionally substituted pyrrolidine, an optionally substitutedpiperidine, an optionally substituted morpholine, or an optionallysubstituted piperazine. In some embodiments, R⁷ is hydrogen and R⁸ isoptionally substituted alkyl, optionally substituted cycloalkyl, oroptionally substituted phenyl. In certain embodiments, R⁷ is hydrogenand R⁸ is selected from methyl, ethyl, propyl, 2-dimethylaminoethyl,2-methoxyethyl, cyclohexyl, and phenyl. In some embodiments, R⁷ and R⁸are hydrogen.

In another aspect, provided herein are compounds of Formula II, orpharmaceutically acceptable salts, polymorphs, solvates, tautomers,metabolites, or N-oxides thereof:

wherein:

-   -   Y¹ and Y² are independently a bond or —N(R⁶)—, wherein at least        one of Y¹ and Y² is —N(R⁶)—;    -   L¹ and L² are independently a bond or optionally substituted        alkylene;    -   X¹ is ═N— or ═C(R²)—;    -   X² is ═N— or ═C(R³)—;    -   R¹¹ is selected from the group consisting of Cl, —CN,        —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹, optionally substituted alkyl,        optionally substituted cycloalkyl, optionally substituted        heterocycloalkyl, optionally substituted alkoxy, haloalkyl,        haloalkoxy, optionally substituted phenyl, and optionally        substituted 5- or 6-membered heteroaryl;    -   R¹⁴ is selected from the group consisting of hydrogen, Cl, Br,        —CN, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹, optionally substituted alkyl,        optionally substituted cycloalkyl, optionally substituted        heterocycloalkyl, optionally substituted alkoxy, haloalkyl,        haloalkoxy, optionally substituted phenyl, and optionally        substituted 5- or 6-membered heteroaryl;    -   R², R³, and R⁵ are independently selected from the group        consisting of hydrogen, halogen, —CN, —C(O)—N(R⁷)—R⁸,        —C(O)—O—R⁹, optionally substituted alkyl, optionally substituted        cycloalkyl, optionally substituted heterocycloalkyl, optionally        substituted alkoxy, haloalkyl, haloalkoxy, optionally        substituted phenyl, and optionally substituted 5- or 6-membered        heteroaryl;    -   R⁶ is hydrogen, optionally substituted alkyl, optionally        substituted alkenyl, or optionally substituted alkynyl;    -   R⁷ and R⁸ are independently hydrogen, optionally substituted        alkyl, haloalkyl, optionally substituted cycloalkyl, optionally        substituted heterocycloalkyl, optionally substituted phenyl, or        R⁷ and R⁸ together with the nitrogen atom to which they are        attached form an optionally substituted heterocycloamino;    -   R⁹ is selected from the group consisting of optionally        substituted alkyl, haloalkyl, optionally substituted cycloalkyl,        optionally substituted heterocycloalkyl, and optionally        substituted phenyl; and    -   A is selected from the group consisting of hydrogen, optionally        substituted alkyl, —OH, optionally substituted alkoxy,        optionally substituted haloalkoxy, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹,        optionally substituted phenyl, and optionally substituted 5- or        6-membered heteroaryl,        -   wherein:            -   if A and R⁵ are hydrogen and R¹ is methoxy, then R⁴ is                independently selected from the group consisting of                hydrogen, —Cl, —CN, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹,                optionally substituted C₂- to C₆-alkyl, optionally                substituted cycloalkyl, optionally substituted                heterocycloalkyl, optionally substituted C₂- to                C₆-alkoxy, haloalkyl, haloalkoxy, optionally substituted                phenyl, and optionally substituted 5- or 6-membered                heteroaryl.

In some embodiments described above or below, provided herein arecompounds of Formula II, wherein Y¹ is a bond and Y² is —N(R⁶)— as shownin Formula IIa:

In certain embodiments described above or below, provided herein arecompounds of Formula II, wherein Y¹ is a bond and Y² is —N(R⁶)—; and X²is ═C(R³)— as shown in Formula IIb:

In some embodiments described above or below, provided herein arecompounds of Formula II, wherein Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; and L¹ is a bond as shown in Formula IIc.

In certain embodiments described above or below, provided herein arecompounds of Formula II, wherein Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; L¹ is a bond; and L² is a bond as shown in Formula IId:

In some embodiments described above or below, provided herein arecompounds of Formula II, wherein Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; L¹ is a bond; L² is a bond; and R⁶ is hydrogen as shown inFormula IIe:

For any embodiments described above or below, provided herein arecompounds of Formula II, wherein L² is an optionally substitutedalkylene.

For any embodiments described above or below, provided herein arecompounds of Formula II, wherein X¹ is ═C(R²)—.

For any embodiments described above or below, provided herein arecompounds of Formula II, wherein R², R³, and R⁵ are independentlyselected from the group consisting of hydrogen, —Cl, —Br, —CN,—C(O)—OMe, methyl, —OMe, and —OCF₃. In some embodiments, R², R³, and R⁵are independently selected from the group consisting of hydrogen, Cl,methyl, and —OMe. In certain embodiments, R² and R³ are hydrogen.

In some embodiments described above or below, provided herein arecompounds of Formula II, wherein R¹¹ is selected from the groupconsisting of —Cl, —CN, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹, methyl, —OMe, —OCF₃,optionally substituted phenyl, and optionally substituted 5- or6-membered heteroaryl; and R¹⁴ is selected from the group consisting ofhydrogen, —Cl, —Br, —CN, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹, methyl, —OMe,—OCF₃, optionally substituted phenyl, and optionally substituted 5- or6-membered heteroaryl. In certain embodiments, R¹⁴ is optionallysubstituted phenyl or optionally substituted 5- or 6-memberedheteroaryl. In some embodiments, R¹⁴ is —C(O)—N(R⁷)—R⁸ or —C(O)—O—R⁹. Incertain embodiments, R¹¹ and R¹⁴ are independently selected from thegroup consisting of —Cl, —Br, —CN, —OMe, and —OCF₃. In some embodiments,R¹¹ is —OMe or —OCF₃.

In certain embodiments described above or below, provided herein arecompounds of Formula II, wherein A is —C(O)—O—R⁹. In some embodiments,R⁹ is hydrogen or methyl.

In some embodiments described above or below, provided herein arecompounds of Formula II, wherein A is —C(O)—N(R⁷)—R⁸. In certainembodiments, R⁷ is hydrogen and R⁸ is optionally substituted alkyl,optionally substituted cycloalkyl, or optionally substituted phenyl. Insome embodiments, R⁷ is hydrogen and R⁸ is selected from hydrogen,methyl, ethyl, propyl, cyclopropyl, and cyclohexyl.

In certain embodiments described above or below, provided herein arecompounds of Formula II, wherein A is selected from optionallysubstituted alkyl, —OH, optionally substituted alkoxy, and optionallysubstituted haloalkoxy. In some embodiments, A is methyl,dimethylaminomethyl, —OH, —OMe, or —OCF₃.

In a further aspect, provided herein are compounds Formula III, orpharmaceutically acceptable salts, polymorphs, solvates, tautomers,metabolites, or N-oxides thereof:

wherein:

-   -   Y¹ and Y² are independently a bond or —N(R⁶)—;    -   L¹ and L² are independently a bond or optionally substituted        alkylene;    -   X¹ is ═N— or ═C(R²)—;    -   X² is ═N— or ═C(R³)—;    -   R¹¹ is selected from the group consisting of Cl, —CN,        —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹, optionally substituted alkyl,        optionally substituted cycloalkyl, optionally substituted        heterocycloalkyl, optionally substituted alkoxy, haloalkyl,        haloalkoxy, optionally substituted phenyl, and optionally        substituted 5- or 6-membered heteroaryl;    -   R¹⁴ is selected from the group consisting of hydrogen, Cl, Br,        —CN, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹, optionally substituted alkyl,        optionally substituted cycloalkyl, optionally substituted        heterocycloalkyl, optionally substituted alkoxy, haloalkyl,        haloalkoxy, optionally substituted phenyl, and optionally        substituted 5- or 6-membered heteroaryl;    -   R², R³, and R⁵ are independently selected from the group        consisting of hydrogen, halogen, —CN, —C(O)—N(R⁷)—R⁸,        —C(O)—O—R⁹, optionally substituted alkyl, optionally substituted        cycloalkyl, optionally substituted heterocycloalkyl, optionally        substituted alkoxy, haloalkyl, haloalkoxy, optionally        substituted phenyl, and optionally substituted 5- or 6-membered        heteroaryl;    -   R⁶ is hydrogen, optionally substituted alkyl, optionally        substituted alkenyl, or optionally substituted alkynyl;    -   R⁷ and R⁸ are independently hydrogen, optionally substituted        alkyl, haloalkyl, optionally substituted cycloalkyl, optionally        substituted heterocycloalkyl, optionally substituted phenyl, or        R⁷ and R⁸ together with the nitrogen atom to which they are        attached form an optionally substituted heterocycloamino;    -   R⁹ is selected from the group consisting of optionally        substituted alkyl, haloalkyl, optionally substituted cycloalkyl,        optionally substituted heterocycloalkyl, and optionally        substituted phenyl; and    -   Z is hydrogen or —N(R¹⁷)—R¹⁸, wherein:        -   if Z and R⁵ are hydrogen and R¹¹ is alkoxy, then R¹⁴ is            independently selected from the group consisting of            hydrogen, Br, —CN, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹, optionally            substituted C₂- to C₆-alkyl, optionally substituted            cycloalkyl, optionally substituted heterocycloalkyl,            optionally substituted C₂- to C₆-alkoxy, haloalkyl,            haloalkoxy, optionally substituted phenyl, and optionally            substituted 5- or 6-membered heteroaryl; and        -   R¹⁷ and R¹⁸ are independently hydrogen, optionally            substituted alkyl, haloalkyl, optionally substituted            cycloalkyl, optionally substituted heterocycloalkyl,            optionally substituted phenyl, or R¹⁷ and R¹⁸ together with            the nitrogen atom to which they are attached form an            optionally substituted heterocycloamino.

In some embodiments described above or below, provided herein arecompounds of Formula III, wherein Y¹ is a bond and Y² is —N(R⁶)— asshown in Formula (IIIa):

In certain embodiments described above or below, provided herein arecompounds of Formula III, wherein Y¹ is a bond and Y² is —N(R⁶)—; and X²is ═C(R³)— as shown in Formula IIIb:

In some embodiments described above or below, provided herein arecompounds of Formula III, wherein Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; and L¹ is a bond as shown in Formula IIIc:

In certain embodiments described above or below, provided herein arecompounds of Formula III, wherein Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; L¹ is a bond; and L² is a bond as shown in Formula IIId:

In some embodiments described above or below, provided herein arecompounds of Formula III, wherein Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; L¹ is a bond; L² is a bond; and R⁶ is hydrogen as shown inFormula IIIe:

For any embodiments described above or below, provided herein arecompounds of Formula III, wherein L² is an optionally substitutedalkylene.

For any embodiments described above or below, provided herein arecompounds of Formula III, wherein X¹ is ═C(R²)—.

For any embodiments described above or below, provided herein arecompounds of Formula III, wherein R², R³, and R⁵ are independentlyselected from the group consisting of hydrogen, —Cl, —Br, —CN,—C(O)—OMe, methyl, —OMe, and —OCF₃. In some embodiments, R², R³, and R⁵are independently selected from the group consisting of hydrogen, Cl,methyl, and —OMe. In certain embodiments, R² and R³ are hydrogen.

In some embodiments provided above or below, provided herein arecompounds of Formula III, wherein R¹¹ is selected from the groupconsisting of Cl, —CN, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹, methyl, —OMe, —OCF₃,optionally substituted phenyl, and optionally substituted 5- or6-membered heteroaryl; and R¹⁴ is selected from the group consisting ofhydrogen, Cl, Br, —CN, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹, methyl, —OMe, —OCF₃,optionally substituted phenyl, and optionally substituted 5- or6-membered heteroaryl. In certain embodiments, R¹⁴ is optionallysubstituted phenyl or optionally substituted 5- or 6-memberedheteroaryl. In some embodiments, R¹⁴ is —C(O)—N(R⁷)—R⁸ or —C(O)—O—R⁹. Incertain embodiments, R¹¹ and R¹⁴ are independently selected from thegroup consisting of —Cl, —Br, —CN, —OMe, and —OCF₃. In some embodiments,R¹¹ is —OMe or —OCF₃.

In certain embodiments provided above or below, provided herein arecompounds of Formula III, wherein Z is —N(R¹⁷)—R¹⁸. In some embodiments,Z is amino, methylamino, dimethylamino, diethylamino,2-methoxyethylamino, dimethylamino, 2-(dimethylamino)ethylamino,morpholine, or 4-methylpiperazinyl.

In another aspect, provided herein are compounds Formula IV, orpharmaceutically acceptable salts, polymorphs, solvates, tautomers,metabolites, or N-oxides thereof:

wherein:

-   -   Y¹ and Y² are independently a bond or —N(R⁶)—;    -   L¹ and L² are independently a bond or optionally substituted        alkylene;    -   X¹ is ═N— or ═C(R²)—;    -   X² is ═N— or ═C(R³)—;    -   R¹ and R⁴ are independently selected from the group consisting        of hydrogen, halogen, —CN, —C(O)—N(R⁷)—R⁸, —C(O)—O—R⁹,        optionally substituted alkyl, optionally substituted cycloalkyl,        optionally substituted heterocycloalkyl, optionally substituted        alkoxy, haloalkyl, haloalkoxy, optionally substituted phenyl,        and optionally substituted 5- or 6-membered heteroaryl;    -   R², R³, and R⁵ are independently selected from the group        consisting of hydrogen, halogen, —CN, —C(O)—N(R⁷)—R⁸,        —C(O)—O—R⁹, optionally substituted alkyl, optionally substituted        cycloalkyl, optionally substituted heterocycloalkyl, optionally        substituted alkoxy, haloalkyl, haloalkoxy, optionally        substituted phenyl, and optionally substituted 5- or 6-membered        heteroaryl;    -   R⁶ is hydrogen, optionally substituted alkyl, optionally        substituted alkenyl, or optionally substituted alkynyl;    -   R⁷ and R⁸ are independently hydrogen, optionally substituted        alkyl, haloalkyl, optionally substituted cycloalkyl, optionally        substituted heterocycloalkyl, optionally substituted phenyl, or        R⁷ and R⁸ together with the nitrogen atom to which they are        attached form an optionally substituted heterocycloamino;    -   R⁹ is selected from the group consisting of hydrogen, optionally        substituted alkyl, haloalkyl, optionally substituted cycloalkyl,        optionally substituted heterocycloalkyl, and optionally        substituted phenyl; and    -   W is selected from the group consisting of an optionally        substituted 5-membered heteroaryl, an optionally substituted        6-membered heteroaryl other than pyridin-3-yl, an optionally        substituted 9-membered heteroaryl, or an optionally substituted        10-membered heteroaryl other than quinolin-3-yl.

In some embodiments described above or below, provided herein arecompounds of Formula IV, wherein Y¹ is a bond and Y² is —N(R⁶)— as shownin Formula IVa:

In certain embodiments described above or below, provided herein arecompounds of Formula IV, wherein Y¹ is a bond and Y² is —N(R⁶)—; and X²is ═C(R³)— as shown in Formula IVb:

In some embodiments described above or below, provided herein arecompounds of Formula IV, wherein Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; and L¹ is a bond as shown in Formula IVc.

In certain embodiments described above or below, provided herein arecompounds of Formula IV, wherein Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; L¹ is a bond; and L² is a bond as shown in Formula IVd:

In some embodiments described above or below, provided herein arecompounds of Formula IV, wherein Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; L¹ is a bond; L² is a bond; and R⁶ is hydrogen as shown inFormula IVe:

For any embodiments described above or below, provided herein arecompounds of Formula IV, wherein L² is an optionally substitutedalkylene.

For any embodiments described above or below, provided herein arecompounds of Formula IV, wherein X¹ is ═C(R²)—.

For any embodiments described above or below, provided herein arecompounds of Formula IV, wherein R², R³, and R⁵ are independentlyselected from the group consisting of hydrogen, —Cl, —Br, —CN,—C(O)—OMe, methyl, —OMe, and —OCF₃. In some embodiments, R², R³, and R⁵are independently selected from the group consisting of hydrogen, Cl,methyl, and —OMe. In certain embodiments, R² and R³ are hydrogen.

In some embodiments described above or below, provided herein arecompounds of Formula IV, wherein R¹ and R⁴ are independently selectedfrom the group consisting of hydrogen, —Cl, —Br, —CN, —C(O)—N(R⁷)—R⁸,—C(O)—O—R⁹, methyl, —OMe, —OCF₃, optionally substituted phenyl, andoptionally substituted 5- or 6-membered heteroaryl. In certainembodiments, R⁴ is optionally substituted phenyl or optionallysubstituted 5- or 6-membered heteroaryl. In some embodiments, R⁴ is—C(O)—N(R⁷)—R⁸ or —C(O)—O—R⁹. In certain embodiments, R¹ and R⁴ areindependently selected from the group consisting of —Cl, —Br, —CN, —OMe,and —OCF₃. In some embodiments, R¹ is —OMe or —OCF₃.

In certain embodiments described above or below, provided herein arecompounds of Formula IV, wherein W is selected from:

wherein:

-   -   R²⁰ is selected from the group consisting of hydrogen, halogen,        —CN, —OH, —C(O)—N(R¹⁷)—R¹⁸, —C(O)—O—R¹⁹, optionally substituted        alkyl, optionally substituted cycloalkyl, optionally substituted        heterocycloalkyl, optionally substituted alkoxy, haloalkyl,        haloalkoxy, optionally substituted phenyl, and optionally        substituted 5- or 6-membered heteroaryl,        -   wherein:            -   R¹⁷ and R¹⁸ are independently hydrogen, optionally                substituted alkyl, haloalkyl, optionally substituted                cycloalkyl, optionally substituted heterocycloalkyl,                optionally substituted phenyl, or R¹⁷ and R¹⁸ together                with the nitrogen atom to which they are attached form                an optionally substituted heterocycloamino; and            -   R¹⁹ is selected from the group consisting of hydrogen,                optionally substituted alkyl, haloalkyl, optionally                substituted cycloalkyl, optionally substituted                heterocycloalkyl, and optionally substituted phenyl; and    -   R²¹ is hydrogen or optionally substituted alkyl.

In some embodiments described above or below, provided herein is acompound of Formula IV, wherein R²⁰ is selected from hydrogen,optionally substituted alkyl, and haloalkyl. In certain embodiments, R²⁰is methyl or trifluoromethyl. In some embodiments, R²¹ is methyl.

Preparation of Compounds

Described herein are compounds of Formula I-IV that inhibit the activityof TNAP, and processes for their preparation. Also described herein arepharmaceutically acceptable salts, pharmaceutically acceptable solvates,pharmaceutically active metabolites, and pharmaceutically acceptableprodrugs of such compounds. Pharmaceutical compositions comprising atleast one such compound or a pharmaceutically acceptable salt,pharmaceutically acceptable solvate, pharmaceutically active metaboliteor pharmaceutically acceptable prodrug of such compound, and apharmaceutically acceptable excipient are also provided.

Compounds of of Formula I-IV may be synthesized using standard syntheticreactions known to those of skill in the art or using methods known inthe art. The reactions can be employed in a linear sequence to providethe compounds or they may be used to synthesize fragments which aresubsequently joined by the methods known in the art.

The starting material used for the synthesis of the compounds describedherein may be synthesized or can be obtained from commercial sources,such as, but not limited to, Aldrich Chemical Co. (Milwaukee, Wis.),Bachem (Torrance, Calif.), or Sigma Chemical Co. (St. Louis, Mo.). Thecompounds described herein, and other related compounds having differentsubstituents can be synthesized using techniques and materials known tothose of skill in the art, such as described, for example, in March,ADVANCED ORGANIC CHEMISTRY 4^(th) Ed., (Wiley 1992); Carey and Sundberg,ADVANCED ORGANIC CHEMISTRY 4^(th) Ed., Vols. A and B (Plenum 2000,2001); Green and Wuts, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS 3^(rd)Ed., (Wiley 1999); Fieser and Fieser's Reagents for Organic Synthesis,Volumes 1-17 (John Wiley and Sons, 1991); Rodd's Chemistry of CarbonCompounds, Volumes 1-5 and Supplementals (Elsevier Science Publishers,1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991); andLarock's Comprehensive Organic Transformations (VCH Publishers Inc.,1989). (all of which are incorporated by reference in their entirety).Other methods for the synthesis of compounds described herein may befound in International Patent Publication No. WO 01/01982901, Arnold etal. Bioorganic & Medicinal Chemistry Letters 10 (2000) 2167-2170;Burchat et al. Bioorganic & Medicinal Chemistry Letters 12 (2002)1687-1690. General methods for the preparation of compound as disclosedherein may be derived from known reactions in the field, and thereactions may be modified by the use of appropriate reagents andconditions, as would be recognized by the skilled person, for theintroduction of the various moieties found in the formulae as providedherein.

The products of the reactions may be isolated and purified, if desired,using conventional techniques, including, but not limited to,filtration, distillation, crystallization, chromatography and the like.Such materials may be characterized using conventional means, includingphysical constants and spectral data.

Compounds described herein may be prepared as a single isomer or amixture of isomers.

Further Forms of Compounds Disclosed Herein

Isomers

In some embodiments, the compounds described herein exist as geometricisomers. In some embodiments, the compounds described herein possess oneor more double bonds. The compounds presented herein include all cis,trans, syn, anti, entgegen (E), and zusammen (Z) isomers as well as thecorresponding mixtures thereof. In some situations, compounds exist astautomers. The compounds described herein include all possible tautomerswithin the formulas described herein. In some situations, the compoundsdescribed herein possess one or more chiral centers and each centerexists in the R configuration, or S configuration. The compoundsdescribed herein include all diastereomeric, enantiomeric, and epimericforms as well as the corresponding mixtures thereof. In additionalembodiments of the compounds and methods provided herein, mixtures ofenantiomers and/or diastereoisomers, resulting from a single preparativestep, combination, or interconversion are useful for the applicationsdescribed herein. In some embodiments, the compounds described hereinare prepared as their individual stereoisomers by reacting a racemicmixture of the compound with an optically active resolving agent to forma pair of diastereoisomeric compounds, separating the diastereomers andrecovering the optically pure enantiomers. In some embodiments,dissociable complexes are preferred (e.g., crystalline diastereomericsalts). In some embodiments, the diastereomers have distinct physicalproperties (e.g., melting points, boiling points, solubilities,reactivity, etc.) and are separated by taking advantage of thesedissimilarities. In some embodiments, the diastereomers are separated bychiral chromatography, or preferably, by separation/resolutiontechniques based upon differences in solubility. In some embodiments,the optically pure enantiomer is then recovered, along with theresolving agent, by any practical means that would not result inracemization.

Labeled Compounds

In some embodiments, the compounds described herein exist in theirisotopically-labeled forms. In some embodiments, the methods disclosedherein include methods of treating diseases by administering suchisotopically-labeled compounds. In some embodiments, the methodsdisclosed herein include methods of treating diseases by administeringsuch isotopically-labeled compounds as pharmaceutical compositions.Thus, in some embodiments, the compounds disclosed herein includeisotopically-labeled compounds, which are identical to those recitedherein, but for the fact that one or more atoms are replaced by an atomhaving an atomic mass or mass number different from the atomic mass ormass number usually found in nature. Examples of isotopes that can beincorporated into compounds of the invention include isotopes ofhydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine andchloride, such as ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸0, ¹⁷O, ³¹P, ³²P, ³⁵S, ¹⁸F,and ³⁶Cl, respectively. Compounds described herein, and the metabolites,pharmaceutically acceptable salts, esters, prodrugs, solvate, hydratesor derivatives thereof which contain the aforementioned isotopes and/orother isotopes of other atoms are within the scope of this invention.Certain isotopically-labeled compounds, for example those into whichradioactive isotopes such as ³H and ¹⁴C are incorporated, are useful indrug and/or substrate tissue distribution assays. Tritiated, i.e., ³Hand carbon-14, i.e., ¹⁴C, isotopes are particularly preferred for theirease of preparation and detectability. Further, substitution with heavyisotopes such as deuterium, i.e., ²H, produces certain therapeuticadvantages resulting from greater metabolic stability, for exampleincreased in vivo half-life or reduced dosage requirements. In someembodiments, the isotopically labeled compounds, pharmaceuticallyacceptable salt, ester, prodrug, solvate, hydrate or derivative thereofis prepared by any suitable method.

In some embodiments, the compounds described herein are labeled by othermeans, including, but not limited to, the use of chromophores orfluorescent moieties, bioluminescent labels, or chemiluminescent labels.

Pharmaceutically Acceptable Salts

In some embodiments, the compounds described herein exist as theirpharmaceutically acceptable salts. In some embodiments, the methodsdisclosed herein include methods of treating diseases by administeringsuch pharmaceutically acceptable salts. In some embodiments, the methodsdisclosed herein include methods of treating diseases by administeringsuch pharmaceutically acceptable salts as pharmaceutical compositions.

In some embodiments, the compounds described herein possess acidic orbasic groups and therefore react with any of a number of inorganic ororganic bases, and inorganic and organic acids, to form apharmaceutically acceptable salt. In some embodiments, these salts areprepared in situ during the final isolation and purification of thecompounds of the invention, or by separately reacting a purifiedcompound in its free form with a suitable acid or base, and isolatingthe salt thus formed.

Examples of pharmaceutically acceptable salts include those saltsprepared by reaction of the compounds described herein with a mineral,organic acid or inorganic base, such salts including, acetate, acrylate,adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate,bisulfite, bromide, butyrate, butyn-1,4-dioate, camphorate,camphorsulfonate, caproate, caprylate, chlorobenzoate, chloride,citrate, cyclopentanepropionate, decanoate, digluconate,dihydrogenphosphate, dinitrobenzoate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptanoate, glycerophosphate, glycolate,hemisulfate, heptanoate, hexanoate, hexyne-1,6-dioate, hydroxybenzoate,γ-hydroxybutyrate, hydrochloride, hydrobromide, hydroiodide,2-hydroxyethanesulfonate, iodide, isobutyrate, lactate, maleate,malonate, methanesulfonate, mandelate metaphosphate, methanesulfonate,methoxybenzoate, methylbenzoate, monohydrogenphosphate,1-napthalenesulfonate, 2-napthalenesulfonate, nicotinate, nitrate,palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, pyrosulfate, pyrophosphate, propiolate, phthalate,phenylacetate, phenylbutyrate, propanesulfonate, salicylate, succinate,sulfate, sulfite, succinate, suberate, sebacate, sulfonate, tartrate,thiocyanate, tosylate undeconate and xylenesulfonate.

Further, the compounds described herein can be prepared aspharmaceutically acceptable salts formed by reacting the free base formof the compound with a pharmaceutically acceptable inorganic or organicacid, including, but not limited to, inorganic acids such ashydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid metaphosphoric acid, and the like; and organic acidssuch as acetic acid, propionic acid, hexanoic acid,cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid,malonic acid, succinic acid, malic acid, maleic acid, fumaric acid,p-toluenesulfonic acid, tartaric acid, trifluoroacetic acid, citricacid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid,mandelic acid, arylsulfonic acid, methanesulfonic acid, ethanesulfonicacid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid,benzenesulfonic acid, 2-naphthalenesulfonic acid,4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylic acid, glucoheptonic acid,4,4′-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionicacid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuricacid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylicacid, stearic acid and muconic acid. In some embodiments, other acids,such as oxalic, while not in themselves pharmaceutically acceptable, areemployed in the preparation of salts useful as intermediates inobtaining the compounds of the invention and their pharmaceuticallyacceptable acid addition salts.

In some embodiments, those compounds described herein which comprise afree acid group react with a suitable base, such as the hydroxide,carbonate, bicarbonate, sulfate, of a pharmaceutically acceptable metalcation, with ammonia, or with a pharmaceutically acceptable organicprimary, secondary, tertiary, or quaternary amine. Representative saltsinclude the alkali or alkaline earth salts, like lithium, sodium,potassium, calcium, and magnesium, and aluminum salts and the like.Illustrative examples of bases include sodium hydroxide, potassiumhydroxide, choline hydroxide, sodium carbonate, N⁺(C₁₋₄ alkyl)₄, and thelike.

Representative organic amines useful for the formation of base additionsalts include ethylamine, diethylamine, ethylenediamine, ethanolamine,diethanolamine, piperazine and the like. It should be understood thatthe compounds described herein also include the quaternization of anybasic nitrogen-containing groups they contain. In some embodiments,water or oil-soluble or dispersible products are obtained by suchquaternization.

Solvates

In some embodiments, the compounds described herein exist as solvates.The invention provides for methods of treating diseases by administeringsuch solvates. The invention further provides for methods of treatingdiseases by administering such solvates as pharmaceutical compositions.

Solvates contain either stoichiometric or non-stoichiometric amounts ofa solvent, and, in some embodiments, are formed during the process ofcrystallization with pharmaceutically acceptable solvents such as water,ethanol, and the like. Hydrates are formed when the solvent is water, oralcoholates are formed when the solvent is alcohol. Solvates of thecompounds described herein can be conveniently prepared or formed duringthe processes described herein. By way of example only, hydrates of thecompounds described herein can be conveniently prepared byrecrystallization from an aqueous/organic solvent mixture, using organicsolvents including, but not limited to, dioxane, tetrahydrofuran ormethanol. In addition, the compounds provided herein can exist inunsolvated as well as solvated forms. In general, the solvated forms areconsidered equivalent to the unsolvated forms for the purposes of thecompounds and methods provided herein.

Polymorphs

In some embodiments, the compounds described herein exist as polymorphs.The invention provides for methods of treating diseases by administeringsuch polymorphs. The invention further provides for methods of treatingdiseases by administering such polymorphs as pharmaceuticalcompositions.

Thus, the compounds described herein include all their crystallineforms, known as polymorphs. Polymorphs include the different crystalpacking arrangements of the same elemental composition of a compound. Incertain instances, polymorphs have different X-ray diffraction patterns,infrared spectra, melting points, density, hardness, crystal shape,optical and electrical properties, stability, and solubility. In certaininstances, various factors such as the recrystallization solvent, rateof crystallization, and storage temperature cause a single crystal formto dominate.

Prodrugs

In some embodiments, the compounds described herein exist in prodrugform. The invention provides for methods of treating diseases byadministering such prodrugs. The invention further provides for methodsof treating diseases by administering such prodrugs as pharmaceuticalcompositions.

Prodrugs are generally drug precursors that, following administration toan individual and subsequent absorption, are converted to an active, ora more active species via some process, such as conversion by ametabolic pathway. Some prodrugs have a chemical group present on theprodrug that renders it less active and/or confers solubility or someother property to the drug. Once the chemical group has been cleavedand/or modified from the prodrug the active drug is generated. Prodrugsare often useful because, in some situations, they are easier toadminister than the parent drug. They are, for instance, bioavailable byoral administration whereas the parent is not. In certain instances, theprodrug also has improved solubility in pharmaceutical compositions overthe parent drug. An example, without limitation, of a prodrug would be acompound as described herein which is administered as an ester (the“prodrug”) to facilitate transmittal across a cell membrane where watersolubility is detrimental to mobility but which then is metabolicallyhydrolyzed to the carboxylic acid, the active entity, once inside thecell where water-solubility is beneficial. A further example of aprodrug might be a short peptide (polyamino acid) bonded to an acidgroup where the peptide is metabolized to reveal the active moiety. (Seefor example Bundgaard, “Design and Application of Prodrugs” in ATextbook of Drug Design and Development, Krosgaard-Larsen and Bundgaard,Ed., 1991, Chapter 5, 113-191, which is incorporated herein byreference).

In some embodiments, prodrugs are designed as reversible drugderivatives, for use as modifiers to enhance drug transport tosite-specific tissues. The design of prodrugs to date has been toincrease the effective water solubility of the therapeutic compound fortargeting to regions where water is the principal solvent.

Additionally, prodrug derivatives of compounds described herein can beprepared by methods described herein are otherwise known in the art (forfurther details see Saulnier et al., Bioorganic and Medicinal ChemistryLetters, 1994, 4, 1985). By way of example only, appropriate prodrugscan be prepared by reacting a non-derivatized compound with a suitablecarbamylating agent, such as, but not limited to,1,1-acyloxyalkylcarbanochloridate, para-nitrophenyl carbonate, or thelike. Prodrug forms of the herein described compounds, wherein theprodrug is metabolized in vivo to produce a derivative as set forthherein are included within the scope of the claims. Indeed, some of theherein-described compounds are prodrugs for another derivative or activecompound.

In some embodiments, prodrugs include compounds wherein an amino acidresidue, or a polypeptide chain of two or more (e.g., two, three orfour) amino acid residues is covalently joined through an amide or esterbond to a free amino, hydroxy or carboxylic acid group of compounds ofthe present invention. The amino acid residues include but are notlimited to the 20 naturally occurring amino acids and also includes4-hydroxyproline, hydroxylysine, demosine, isodemosine,3-methylhistidine, norvaline, beta-alanine, gamma-aminobutyric acid,cirtulline, homocysteine, homoserine, ornithine and methionine sulfone.In other embodiments, prodrugs include compounds wherein a nucleic acidresidue, or an oligonucleotide of two or more (e.g., two, three or four)nucleic acid residues is covalently joined to a compound of the presentinvention.

Pharmaceutically acceptable prodrugs of the compounds described hereinalso include, but are not limited to, esters, carbonates,thiocarbonates, N-acyl derivatives, N-acyloxyalkyl derivatives,quaternary derivatives of tertiary amines, N-Mannich bases, Schiffbases, amino acid conjugates, phosphate esters, metal salts andsulfonate esters. Compounds having free amino, amido, hydroxy orcarboxylic groups can be converted into prodrugs. For instance, freecarboxyl groups can be derivatized as amides or alkyl esters. In certaininstances, all of these prodrug moieties incorporate groups includingbut not limited to ether, amine and carboxylic acid functionalities.

Hydroxy prodrugs include esters, such as though not limited to,acyloxyalkyl (e.g. acyloxymethyl, acyloxyethyl) esters,alkoxycarbonyloxyalkyl esters, alkyl esters, aryl esters, phosphateesters, sulfonate esters, sulfate esters and disulfide containingesters; ethers, amides, carbamates, hemisuccinates,dimethylaminoacetates and phosphoryloxymethyloxycarbonyls, as outlinedin Advanced Drug Delivery Reviews 1996, 19, 115.

Amine derived prodrugs include, but are not limited to the followinggroups and combinations of groups:

as well as sulfonamides and phosphonamides.

In certain instances, sites on any aromatic ring portions aresusceptible to various metabolic reactions, therefore incorporation ofappropriate substituents on the aromatic ring structures, can reduce,minimize or eliminate this metabolic pathway.

Metabolites

In some embodiments, compounds of Formula I-IV are susceptible tovarious metabolic reactions Therefore, in some embodiments,incorporation of appropriate substituents into the structure willreduce, minimize, or eliminate a metabolic pathway. In specificembodiments, the appropriate substituent to decrease or eliminate thesusceptibility of an aromatic ring to metabolic reactions is, by way ofexample only, a halogen, or an alkyl group.

In additional or further embodiments, the compounds of Formula I-IVdescribed herein are metabolized upon administration to an organism inneed to produce a metabolite that is then used to produce a desiredeffect, including a desired therapeutic effect.

Pharmaceutical Compositions/Formulations

In a further aspect provided herein are pharmaceutical compositionscomprising a compound of Formula I, Formula II, Formula III, or FormulaIV, or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable excipient.

In some embodiments, the compounds described herein are formulated intopharmaceutical compositions. Pharmaceutical compositions are formulatedin a conventional manner using one or more pharmaceutically acceptableinactive ingredients that facilitate processing of the active compoundsinto preparations that can be used pharmaceutically. Proper formulationis dependent upon the route of administration chosen. A summary ofpharmaceutical compositions described herein can be found, for example,in Remington: The Science and Practice of Pharmacy, Nineteenth Ed(Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E.,Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa.1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical DosageForms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical DosageForms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams &Wilkins 1999), herein incorporated by reference for such disclosure.

Provided herein are pharmaceutical compositions that include a compoundof Formula I-IV and at least one pharmaceutically acceptable inactiveingredient. In some embodiments, the compounds described herein areadministered as pharmaceutical compositions in which a compound ofFormula I-IV is mixed with other active ingredients, as in combinationtherapy. In other embodiments, the pharmaceutical compositions includeother medicinal or pharmaceutical agents, carriers, adjuvants,preserving, stabilizing, wetting or emulsifying agents, solutionpromoters, salts for regulating the osmotic pressure, and/or buffers. Inyet other embodiments, the pharmaceutical compositions include othertherapeutically valuable substances.

A pharmaceutical composition, as used herein, refers to a mixture of acompound of Formula I-IV with other chemical components (i.e.pharmaceutically acceptable inactive ingredients), such as carriers,excipients, binders, filling agents, suspending agents, flavoringagents, sweetening agents, disintegrating agents, dispersing agents,surfactants, lubricants, colorants, diluents, solubilizers, moisteningagents, plasticizers, stabilizers, penetration enhancers, wettingagents, anti-foaming agents, antioxidants, preservatives, or one or morecombination thereof. The pharmaceutical composition facilitatesadministration of the compound to an organism. In practicing the methodsof treatment or use provided herein, therapeutically effective amountsof compounds described herein are administered in a pharmaceuticalcomposition to a mammal having a disease, disorder, or condition to betreated. In some embodiments, the mammal is a human. A therapeuticallyeffective amount can vary widely depending on the severity of thedisease, the age and relative health of the subject, the potency of thecompound used and other factors. The compounds can be used singly or incombination with one or more therapeutic agents as components ofmixtures.

The pharmaceutical formulations described herein are administered to asubject by appropriate administration routes, including but not limitedto, oral, parenteral (e.g., intravenous, subcutaneous, intramuscular),intranasal, buccal, topical, rectal, or transdermal administrationroutes. The pharmaceutical formulations described herein include, butare not limited to, aqueous liquid dispersions, liquids, gels, syrups,elixirs, slurries, suspensions, self-emulsifying dispersions, solidsolutions, liposomal dispersions, aerosols, solid oral dosage forms,powders, immediate release formulations, controlled releaseformulations, fast melt formulations, tablets, capsules, pills, powders,dragees, effervescent formulations, lyophilized formulations, delayedrelease formulations, extended release formulations, pulsatile releaseformulations, multiparticulate formulations, and mixed immediate andcontrolled release formulations.

Pharmaceutical compositions including a compound of Formula I-IV aremanufactured in a conventional manner, such as, by way of example only,by means of conventional mixing, dissolving, granulating, dragee-making,levigating, emulsifying, encapsulating, entrapping or compressionprocesses.

The pharmaceutical compositions will include at least one compound ofFormula I-IV as an active ingredient in free-acid or free-base form, orin a pharmaceutically acceptable salt form. In addition, the methods andpharmaceutical compositions described herein include the use of N-oxides(if appropriate), crystalline forms, amorphous phases, as well as activemetabolites of these compounds having the same type of activity. In someembodiments, compounds described herein exist in unsolvated form or insolvated forms with pharmaceutically acceptable solvents such as water,ethanol, and the like. The solvated forms of the compounds presentedherein are also considered to be disclosed herein.

Pharmaceutical preparations for oral use are obtained by mixing one ormore solid excipient with one or more of the compounds described herein,optionally grinding the resulting mixture, and processing the mixture ofgranules, after adding suitable auxiliaries, if desired, to obtaintablets or dragee cores. Suitable excipients include, for example,fillers such as sugars, including lactose, sucrose, mannitol, orsorbitol; cellulose preparations such as, for example, maize starch,wheat starch, rice starch, potato starch, gelatin, gum tragacanth,methylcellulose, microcrystalline cellulose,hydroxypropylmethylcellulose, sodium carboxymethylcellulose; or otherssuch as: polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. Ifdesired, disintegrating agents are added, such as the cross-linkedcroscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid or asalt thereof such as sodium alginate. In some embodiments, dyestuffs orpigments are added to the tablets or dragee coatings for identificationor to characterize different combinations of active compound doses.

Pharmaceutical preparations that are administered orally includepush-fit capsules made of gelatin, as well as soft, sealed capsules madeof gelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules contain the active ingredients in admixture with filler such aslactose, binders such as starches, and/or lubricants such as talc ormagnesium stearate and, optionally, stabilizers. In soft capsules, theactive compounds are dissolved or suspended in suitable liquids, such asfatty oils, liquid paraffin, or liquid polyethylene glycols. In someembodiments, stabilizers are added.

In certain embodiments, delivery systems for pharmaceutical compoundsmay be employed, such as, for example, liposomes and emulsions. Incertain embodiments, compositions provided herein can also include anmucoadhesive polymer, selected from among, for example,carboxymethylcellulose, carbomer (acrylic acid polymer),poly(methylmethacrylate), polyacrylamide, polycarbophil, acrylicacid/butyl acrylate copolymer, sodium alginate and dextran.

Methods

In another aspect provided herein are methods of treating a disease in asubject mediated by tissue-nonspecific alkaline phosphatase (TNAP),which method comprises administering to the subject a pharmaceuticalcomposition comprising a compound of Formula I, Formula II, Formula III,or Formula IV, or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable excipient. In some embodiments, the diseaseis a medial vascular calcification, ectopic ossification in spinalligaments, ankylosis, or osteoarthritis. In certain embodiments, thedisease is arterial calcification.

In some embodiments, administration of a therapeutically effectiveamount of a compound of Formula I-IV retards or reverses the formation,growth or deposition of extracellular matrix hydroxyapatite crystaldeposits. In certain embodiments, administration of a compound ofFormula I-IV retards or reverses the formation, growth or deposition ofextracellular matrix hydroxyapatite crystal deposits. In someembodiments, administration of the compound of the invention preventsthe formation, growth or deposition of extracellular matrixhydroxyapatite crystal deposits.

In some embodiments, the invention provides a method of inhibiting,decreasing, treating, or preventing pathological calcification in anindividual by administration of a therapeutically effective amount of acompound of Formula I-IV. In some embodiments, the present inventionprovides a method of treating or preventing a disorder characterized bypathological calcification, such as ankylosing spondylitis, tumoralcalcinosis, fibrodysplasia ossificans progressiva, progressive osseousheteroplasia, pseudoxanthoma elasticum, ankylosis, osteoarthritis,general arterial calcification in infancy (GACI), arterial calcificationdue to deficiency of CD73 (ACDC), Keutel syndrome, peritonealcalcification, heterotopic calcification/ossification in amputees,tibial artery calcification, bone metastasis, prosthetic calcification,and/or Paget's disease of bone.

In certain embodiments, the invention provides a method of inhibiting,decreasing, treating, or preventing vascular calcification in anindividual by administration of a compound of Formula I-IV. In someembodiments, the present invention provides a method of treating orpreventing atherosclerotic calcification, medial calcification and otherconditions characterized by vascular calcification. In some embodiments,the present invention provides a method of treating or preventingvascular calcification associated with diabetes mellitus I and II,idiopathic infantile arterial calcification (IIAC), Kawasaki disease,obesity, and/or increased age.

In some embodiments, the invention provides a method of inhibiting,decreasing, treating, or preventing vascular calcification associatedwith chronic renal disease (chronic renal insufficiency) or end-stagerenal disease by administration of a therapeutically effective amount ofa compound of Formula I-IV. In some embodiments, the invention providesa method of inhibiting, decreasing or preventing vascular calcificationassociated with pre- or post-dialysis uremia. In some embodiments, theinvention provides a method of inhibiting, decreasing or preventingvascular calcification-associated chronic kidney disease, wherein thechronic kidney disease is associated with secondary hyperparathyroidism(HPT) characterized by elevated parathyroid hormone (PTH) levels. Insome embodiments, the invention provides a method of inhibiting,decreasing or preventing symptoms associated with calciphylaxis, orcalcific uremic arteriolopathy. In some embodiments, the inventionprovides a method of administering an effective amount of a TNAPinhibitor for reducing serum PTH without causing aortic calcification.In some embodiments, the invention provides a method of administering aneffective amount of a TNAP inhibitor (e.g., a compound of Formula I-IV)for reducing serum creatinine level or preventing increase of serumcreatinine level.

Assessment of Vascular Calcification

Methods of detecting and measuring vascular calcification are well knownin the art. In some embodiments, methods of measuring calcificationinclude direct methods of detecting and measuring extent ofcalcium-phosphorus depositions in blood vessels.

In some embodiments, direct methods of measuring vascular calcificationcomprise in vivo imaging methods such as plain film roentgenography,coronary arteriography; fluoroscopy, including digital subtractionfluoroscopy; cinefluorography; conventional, helical, and electron beamcomputed tomography; intravascular ultrasound (IVUS); magnetic resonanceimaging; and transthoracic and transesophageal echocardiography. Personsskilled in the art most commonly use fluoroscopy and EBCT to detectcalcification noninvasively. Coronary interventionalists usecinefluorography and IVUS to evaluate calcification in specific lesionsbefore angioplasty.

In some embodiments, vascular calcification can be detected by plainfilm roentgenography. The advantage of this method is availability ofthe film and the low cost of the method, however, the disadvantage isits low sensitivity (Kelley M. & Newell J. Cardiol Clin. 1: 575-595,1983). In some embodiments, fluoroscopy can be used to detectcalcification in coronary arteries. Although fluoroscopy can detectmoderate to large calcifications, its ability to identify small calcificdeposits is low (Loecker et al. J. Am. Coll. Cardiol. 19: 1167-1172,1992). Fluoroscopy is widely available in both inpatient and outpatientsettings and is relatively inexpensive. In some embodiments, vasculardetection can be detected by conventional computed tomography (CT).Because calcium attenuates the x-ray beam, computed tomography (CT) isextremely sensitive in detecting vascular calcification. Whileconventional CT appears to have better capability than fluoroscopy todetect coronary artery calcification, its limitations are slow scantimes resulting in motion artifacts, volume averaging, breathingmisregistration, and inability to quantify amount of plaque (Wexler etal. Circulation 94: 1175-1192, 1996).

In some embodiments, calcification can be detected by helical or spiralcomputer tomography, which has considerably faster scan times thanconventional CT. Overlapping sections also improve calcium detection.Coronary calcium imaging by helical CT has a sensitivity of 91% and aspecificity of 52% when compared with angiographically significantcoronary obstructive disease (Shemesh et al. Radiology 197: 779-783,1995). Double-helix CT scanners appear to be more sensitive thansingle-helix scanners in detection of coronary calcification because oftheir higher resolution and thinner slice capabilities.

In some embodiments, Electron Beam Computed Tomography (EBCT) can beused for detection of vascular calcification. EBCT uses an electron gunand a stationary tungsten “target” rather than a standard x-ray tube togenerate x-rays, permitting very rapid scanning times. Originallyreferred to as cine or ultrafast CT, the term EBCT is now used todistinguish it from standard CT scans because modern spiral scanners arealso achieving subsecond scanning times. For purposes of detectingcoronary calcium, EBCT images are obtained in 100 ms with a scan slicethickness of 3 mm. Thirty to 40 adjacent axial scans are obtained bytable incrementation. The scans, which are usually acquired during oneor two separate breath-holding sequences, are triggered by theelectrocardiographic signal at 80% of the RR interval, near the end ofdiastole and before atrial contraction, to minimize the effect ofcardiac motion. The rapid image acquisition time virtually eliminatesmotion artifact related to cardiac contraction. The unopacified coronaryarteries are easily identified by EBCT because the lower CT density ofperiarterial fat produces marked contrast to blood in the coronaryarteries, while the mural calcium is evident because of its high CTdensity relative to blood. Additionally, the scanner software allowsquantification of calcium area and density. An arbitrary scoring systemhas been devised based on the x-ray attenuation coefficient, or CTnumber measured in Hounsfield units, and the area of calcified deposits(Agatston et al. J. Am. Coll. Cardiol. 15:827-832, 1990). A screeningstudy for coronary calcium can be completed within 10 or 15 minutes,requiring only a few seconds of scanning time. Electron beam CT scannersare more expensive than conventional or spiral CT scanners and areavailable in relatively fewer sites.

In some embodiments, intravascular ultrasound (IVUS) can be used fordetecting vascular calcification, in particular, coronaryatherosclerosis (Waller et al. Circulation 85: 2305-2310, 1992). Byusing transducers with rotating reflectors mounted on the tips ofcatheters, it is possible to obtain cross-sectional images of thecoronary arteries during cardiac catheterization. The sonograms provideinformation not only about the lumen of the artery but also about thethickness and tissue characteristics of the arterial wall. Calcificationis seen as a hyperechoic area with shadowing: fibrotic noncalcifiedplaques are seen as hyperechoic areas without shadowing (Honye et al.Trends Cardiovasc Med. 1: 305-311, 1991). The disadvantages in use ofIVUS, as opposed to other imaging modalities, are that it is invasiveand currently performed only in conjunction with selective coronaryangiography, and it visualizes only a limited portion of the coronarytree. Although invasive, the technique is clinically important becauseit can show atherosclerotic involvement in patients with normal findingson coronary arteriograms and helps define the morphologicalcharacteristics of stenotic lesions before balloon angioplasty andselection of atherectomy devices (Tuzcu et al. J. Am. Coll. Cardiol. 27:832-838, 1996).

In some embodiments, vascular calcification can be measured by magneticresonance imaging (MRI). In some embodiments, vascular calcification canbe measured by transthoracic (surface) echocardiography, which isparticularly sensitive to detection of mitral and aortic valvularcalcification. In some embodiments, vascular calcification can beassessed ex vivo by Van Kossa method. This method relies upon theprinciple that silver ions can be displaced from solution by carbonateor phosphate ions due to their respective positions in theelectrochemical series. The argentaffin reaction is photochemical innature and the activation energy is supplied from strong visible orultra-violet light. Since the demonstrable forms of tissue carbonate orphosphate ions are invariably associated with calcium ions the methodcan be considered as demonstrating sites of tissue calcium deposition.

Other methods of direct measuring calcification may include, but notlimited to, immuno fluorescent staining and densitometry. In anotheraspect, methods of assessing vascular calcification include methods ofmeasuring determinants and/or risk factors of vascular calcification.Such factors include, but are not limited to, serum levels ofphosphorus, calcium, and calcium phosphorus product, parathyroid hormone(PTH), low-density lipoprotein cholesterol (LDL), high-densitylipoprotein cholesterol (HDL), triglycerides, and creatinine. Methods ofmeasuring these factors are well known in the art. Other methods ofassessing vascular calcification include assessing factors of boneformation. Such factors include bone formation markers such asbone-specific alkaline phosphatase (BSAP), osteocalcin (OC),carboxyterminal propeptide of type I collagen (PICP), and aminoterminalpropeptide of type I collagen (PINP); serum bone resorption markers suchas cross-linked C-telopeptide of type I collagen (ICTP),tartrate-resistant acid phosphatase, TRACP and TRAP5B, N-telopeptide ofcollagen cross-links (NTx), and C-telopeptide of collagen cross-links(CTx); and urine bone resorption markers, such as hydroxyproline, freeand total pyridinolines (Pyd), free and total deoxypyridinolines (Dpd),N-telopeptide of collagen cross-links (NTx), and C-telopeptide ofcollagen crosslinks (CTx).

Administration of Pharmaceutical Composition

Suitable routes of administration include, but are not limited to, oral,intravenous, rectal, aerosol, parenteral, ophthalmic, pulmonary,transmucosal, transdermal, vaginal, otic, nasal, and topicaladministration. In addition, by way of example only, parenteral deliveryincludes intramuscular, subcutaneous, intravenous, intramedullaryinjections, as well as intrathecal, direct intraventricular,intraperitoneal, intralymphatic, and intranasal injections.

In certain embodiments, a compound as described herein is administeredin a local rather than systemic manner, for example, via injection ofthe compound directly into an organ, often in a depot preparation orsustained release formulation. In specific embodiments, long actingformulations are administered by implantation (for examplesubcutaneously or intramuscularly) or by intramuscular injection.Furthermore, in other embodiments, the drug is delivered in a targeteddrug delivery system, for example, in a liposome coated withorgan-specific antibody. In such embodiments, the liposomes are targetedto and taken up selectively by the organ. In yet other embodiments, thecompound as described herein is provided in the form of a rapid releaseformulation, in the form of an extended release formulation, or in theform of an intermediate release formulation. In yet other embodiments,the compound described herein is administered topically.

In some embodiments, TNAP inhibitors (e.g., a compound of Formula I-IV)is administered alone or in combination with other drugs for treatingvascular calcification, such as vitamin D sterols and/or RENAGEL®.Vitamin D sterols can include calcitriol, alfacalcidol, doxercalciferol,maxacalcitol or paricalcitol. In certain embodiments, the compounds ofFormula I-IV are used with calcimimetics, vitamins and their analogs,antibiotics, lanthanum carbonate, lipid-lowering agents, such asLIPITOR®, anti-hypertensives, anti-inflammatory agents (steroidal andnon-steroidal), inhibitors of pro-inflammatory cytokine (ENBREL®,KINERET®), and cardiovascular agents.

In some embodiments, the compositions disclosed herein are administeredbefore, concurrently, or after administration of calcimimetics, vitaminD sterols and/or RENAGEL®. The dosage regimen for treating a diseasecondition with the combination therapy disclosed herein is selected inaccordance with a variety of factors, including the type, age, weight,sex and medical condition of the patient, the severity of the disease,the route of administration, and the particular compound employed, andthus can vary widely.

In some embodiments, TNAP inhibitors (e.g., compound of Formula I-IV)are administered before or after administration of vitamin D sterols. Insome embodiments, TNAP inhibitors are coadministered with vitamin Dsterols. In certain embodiments, the methods disclosed herein arepracticed to attenuate the mineralizing effect of calcitriol on vasculartissue. In some embodiments, the methods disclosed herein are used toreverse the effect of calcitriol of increasing the serum levels ofcalcium, phosphorus and calcium-phosphorus product thereby preventing orinhibiting vascular calcification. In some embodiments, the methodsdisclosed herein are used to stabilize or decrease serum creatininelevels. In some embodiments, in addition to creatinine level increasedue to a disease, a further increase in creatinine level is due totreatment with vitamin D sterols such as calcitriol.

In additional embodiments, the compounds of Formula I-IV areadministered in conjunction with surgical and non-surgical treatments.In one aspect, the methods disclosed herein can be practiced ininjunction with dialysis.

In some embodiments, compounds of Formula I-IV and compositions thereofare administered in any suitable manner. The manner of administrationcan be chosen based on, for example, whether local or systemic treatmentis desired, and on the area to be treated. For example, the compositionscan be administered orally, parenterally (e.g., intravenous,subcutaneous, intraperitoneal, or intramuscular injection), byinhalation, extracorporeally, topically (including transdermally,ophthalmically, vaginally, rectally, intranasally) or the like. In someembodiments, a compound of Formula I-IV is administered directly to thesite of hyper-mineralization using a drug delivery device orformulation. In certain embodiments, the drug delivery device orformulation releases the compound for Formula I-IV over a period of timeat the local target site.

It is further understood and herein contemplated that the disclosedinhibitors can be administered in conjunction with balloons tippedcatheters and/or stents. It is contemplated herein that the stents,catheters, and/or balloons can be linked with the TNAP inhibitors (e.g.,compounds of Formula I-IV) or administered concurrently with the use. By“linking” or “linked” is meant any method of placing a TNAP inhibitoronto the stent such as soaking, coating, infusing, or any known chemicalmethods. Also contemplated herein are time released methods of attachinga TNAP inhibitor to a balloon or stent. Thus, for example disclosedherein are stents used for treatment of a vascular condition, whereinthe stent has been coated with a TNAP inhibitor. Also disclosed hereinare methods of inhibiting, decreasing or preventing vascularcalcification comprising administering to an individual a stent,balloon, and/or catheter that has been linked to a TNAP inhibitor. Thus,for example disclosed herein are methods of inhibiting, decreasing orpreventing vascular calcification comprising administering to a subjecta vascular stent coated with a TNAP inhibitor.

It is further understood and herein contemplated that the disclosedinhibitors can be administered in conjunction with prostheses, such as aprosthetic heart valve. It is contemplated herein that the prosthesiscan be linked with the TNAP inhibitors or administered concurrently withthe use. By “linking” or “linked” is meant any method of placing a TNAPinhibitor onto the prosthesis, such as soaking, coating, infusing, orany known chemical methods. Also contemplated herein are time releasedmethods of attaching a TNAP inhibitor to a prosthesis. Thus, for exampledisclosed herein are prostheses used for treatment of a vascularcondition, wherein the prosthesis has been coated with a TNAP inhibitor.Also disclosed herein are methods of inhibiting, decreasing orpreventing prosthesis calcification, comprising administering to anindividual a prosthesis that has been linked to a TNAP inhibitor.

It is further understood and herein contemplated that the disclosedinhibitors can be administered as a drug implant. It is contemplatedherein that the TNAP inhibitors can be formulated into a sustainedrelease particle for localized tissue insertion. Also contemplatedherein are time released methods of localized administration of TNAP toa particular region of tissue. Thus, for example disclosed herein areTNAP inhibitor drug implants used for treatment of heterotopicossification, wherein the sustained-release TNAP inhibitor drug implantis locally placed at a site of undesired hydroxyapatite deposition, suchas subcutaneously at a site of amputation.

Parenteral administration of the composition, if used, is generallycharacterized by injection. Injectables can be prepared in conventionalforms, either as liquid solutions or suspensions, solid forms suitablefor solution of suspension in liquid prior to injection, or asemulsions. A more recently revised approach for parenteraladministration involves use of a slow release or sustained releasesystem such that a constant dosage is maintained.

EXAMPLES

The following preparations of compound of Formula I-IV and intermediatesare given to enable those of skill in the art to more clearly understandand to practice the present invention. They should not be considered aslimiting the scope of the invention, but merely as illustrative andrepresentative thereof.

SYNTHETIC EXAMPLES Example I Example I-15-Chloro-2-methoxy-N-pyridin-3-yl-benzenesulfonamide

A mixture of 5-chloro-2-methoxy-benzenesulfonyl chloride (213 mg, 0.88mmol), pyridine-3-ylamine (100 mg, 0.88 mmol), DMAP (10 mg, cat.) inpyridine (5 mL) was stirred at 50° C. for 2 h. LCMS indicated thereaction was complete. The solvent was evaporated in vacuum. The residuewas treated with DCM (5 mL). The suspension was collected by filtrationto give crude product, which was purified by prep-HPLC to afford 100 mg(yield: 38%) of 5-chloro-2-methoxy-N-pyridin-3-yl-benzenesulfonamide aspale yellow solid.

¹H NMR (DMSO-d6): δ=10.45 (1H, brs), 8.37 (1H, s), 8.32 (1H, d), 7.74(1H, s), 7.69-7.64 (2H, m), 7.38 (1H, q), 7.25 (1H, d), 3.85 (3H, s).MS: m/z 398.9 (M+H⁺).

Example I-22-Methoxy-N-pyridin-3-yl-5-trifluoromethyl-benzenesulfonamide

Step 1: To chlorosulfuric acid (15 mL) was added1-methoxy-4-trifluoromethyl-benzene (3.0 g, 17 mmol) portionwise at 0°C. The mixture was stirred at room temperature overnight. The mixturewas poured into ice. the aqueous layer was extracted with EtOAc (50mL×3). The extracts were dried over Na₂SO₄ and the solution was filteredthrough a pad of silica gel, dried in vacuum to afford 500 mg (yield:11%) of 2-methoxy-5-trifluoromethyl-benzenesulfonyl chloride as whitesolid.

¹H NMR (DMSO-d6): δ=7.90 (1H, d), 7.67 (1H, dd), 7.18 (1H, d), 3.84 (3H,s).

Step 2: The procedure is similar to Example I-1.

¹H NMR (DMSO-d6): δ=10.70 (1H, brs), 8.35 (1H, d), 8.28 (1H, dd),8.03-7.98 (2H, m), 7.62-7.58 (1H, m), 7.43-7.34 (2H, m), 3.92 (3H, s).MS: m/z 332.9 (M+H⁺).

Example I-3 5-Bromo-2-methoxy-N-pyridin-3-yl-benzenesulfonamide

This compound was prepared as described in Example I-1.

¹H NMR (DMSO-d6): δ=10.45 (1H, brs), 8.30 (1H, d), 8.23 (1H, d),7.78-7.74 (2H, m), 7.49 (1H, d), 7.28 (1H, dd), 7.17 (1H, d), 3.85 (3H,s). MS: m/z 344.8 (M+H⁺).

Example 1-4 2-Methoxy-4-methyl-N-pyridin-3-yl-benzenesulfonamide

This compound was prepared as described in Example I-1.

1H NMR (DMSO-d6): δ=10.80 (1H, brs), 8.34 (1H, s), 8.26 (1H, d), 7.66(1H, d), 7.63-7.60 (1H, m), 7.42-7.36 (1H, m), 7.01 (1H, s), 6.86 (1H,d), 3.82 (3H, s), 2.32 (3H, s). MS: m/z 279.0 (M+H⁺).

Example I-5 2,4-Dimethoxy-N-pyridin-3-yl-benzenesulfonamide

This compound was prepared as described in Example I-1.

¹H NMR (DMSO-d6): δ=10.17 (1H, brs), 8.29 (1H, d), 8.18 (1H, d), 7.69(1H, d), 7.51 (1H, dd), 7.30-7.23 (1H, m), 6.64 (1H, d), 6.58 (1H, d),3.85 (3H, s), 3.79 (3H, s). MS: m/z 295.0 (M+H⁺).

Example I-6 5-Cyano-2-methoxy-N-pyridin-3-yl-benzenesulfonamide

A mixture of 5-bromo-2-methoxy-N-(pyridin-3-yl)benzenesulfonamide (100mg, 0.29 mmol), Zn(CN)₂ (136 mg, 1.16 mmol) and Pd(PPh₃)₄ (5% cat.amount) in DMF (3 mL) was bubbled with N₂ for 5 min and heated at 120°C. for 1 h under microwave irridation. After cooled to room temperature,the solvent was evaporated in vacuum. The residue was partitionedbetween DCM (5 mL) and H₂O (10 mL). The mixture was extracted with DCM(15 mL×3). The extracts were dried over Na₂SO₄ and concentrated invacuum to give crude compound, which was purified by prep-HPLC to afford30 mg (yield: 36%) of5-cyano-2-methoxy-N-pyridin-3-yl-benzenesulfonamide as white solid.

¹H NMR (DMSO-d6): δ=10.57 (1H, brs), 8.30 (1H, s), 8.23 (1H, s), 8.17(1H, s), 8.08 (1H, d), 7.49 (1H, d), 7.38 (1H, d), 7.28 (1H, d), 3.95(s, 3H). MS: m/z 290.0 (M+H⁺).

Example I-7 4-Methoxy-biphenyl-3-sulfonic acid pyridin-3-ylamide

The mixture of 5-bromo-2-methoxy-N-pyridin-3-yl-benzenesulfonamide (50mg, 0.15 mmol), phenylboronic acid (34 mg, 0.29 mmol), Pd(PPh₃)₄ (20 mg,cat.), K₂CO₃ (40 mg, 0.30 mmol) in DMF (2 mL) was stirred at 130° C. for30 min under microwave irridation. After cooled to room temperature, thesolvent was evaporated in vacuum. The residue was partitioned betweenDCM (5 mL) and H₂O (10 mL). The mixture was extracted with DCM (15mL×3). The extracts were dried over Na₂SO₄ and concentrated in vacuum togive crude compound, which was purified by prep-TLC (EtOAc) to afford 22mg (yield: 43%) of 4-methoxy-biphenyl-3-sulfonic acid pyridin-3-ylamideas yellow solid.

¹H NMR (CDCl3): δ=8.32 (1H, d), 8.24 (1H, d), 8.03 (1H, d), 7.73 (1H,dd), 7.66 (1H, d), 7.47 (2H, d), 7.42 (2H, t), 7.34 (1H, t), 7.19 (1H,t), 7.14 (1H, s), 7.08 (1H, s), 4.01 (3H, s). MS: m/z 341.0 (M+H⁺).

Example I-8 2-Methoxy-N-pyridin-3-yl-5-thiophen-3-yl-benzenesulfonamide

¹H NMR (DMSO-d6): δ=10.81 (1H, brs), 8.28 (1H, s), 8.11 (1H, d), 8.03(1H, d), 7.88 (1H, d), 7.83 (1H, s), 7.84 (1H, t), 7.48 (1H, d), 7.46(1H, s), 7.21-7.17 (2H, m), 3.87 (3H, s). MS: m/z 347.0 (M+H⁺).

Example I-9 5-Furan-3-yl-2-methoxy-N-pyridin-3-yl-benzenesulfonamide

¹H NMR (DMSO-d6): δ=10.44 (1H, brs), 8.35 (1H, s), 8.22-8.18 (2H, m),7.94 (1H, s), 7.83 (1H, d), 7.74 (1H, s), 7.57 (1H, d), 7.32 (1H, d),7.22 (1H, d), 6.94 (1H, s), 3.87 (3H, s). MS: m/z 331.0 (M+H⁺).

Example I-10 2-Methoxy-5-pyridin-4-yl-N-pyridin-3-yl-benzenesulfonamide

Step 1: The mixture of 1-bromo-4-methoxy-benzene (1.8 g, 10 mmol),pyridine-4-boronic acid (10 mmol), K₂CO₃ (2.7 g, 20 mmol), Pd(PPh₃)₄(400 mg) in DMF (40 mL) was stirred at 120° C. for 4 h. TLC indicatedthe reaction was complete. The solvent was evaporated in vacuum. Theresidue was purified by silica gel column (PE/EtOAc, 20/1) to afford1.28 g of 4-(4-methoxy-phenyl)-pyridine (yield: 69%) as white solid. MS:m/z 186.0 (M+H⁺).

Step 2: To chlorosulfuric acid (10 mL) was added4-(4-methoxy-phenyl)-pyridine (1.28 g, 6.91 mmol) portionwise at 0° C.and the mixture was stirred at room temperature for 4 h. The mixture waspoured into ice-water, neutralized with saturated NaHCO₃ to pH=7-8 andextracted with EtOAc (25 mL×3). The extracts were dried over Na₂SO₄ andconcentrated in vacuum to afford 600 mg (yield: 31%) of2-methoxy-5-pyridin-4-yl-benzenesulfonyl chloride.

¹H NMR (DMSO-d6): δ=8.70 (1H, s), 8.29 (1H, s), 8.09 (1H, d), 7.77 (1H,d), 7.44-7.41 (2H, m), 7.36 (1H, d), 4.12 (3H, s).

Step 3: The procedure is similar to Example I-1.

¹H NMR (DMSO-d6): δ=10.45 (1H, brs), 8.61 (2H, d), 8.33 (1H, d), 8.19(1H, d), 8.11 (1H, d), 8.07 (1H, dd), 7.67 (2H, d), 7.53 (1H, d), 7.34(1H, d), 7.25 (1H, d), 3.93 (3H, s). MS: m/z 299.0 (M+H⁺).

Example I-11 4-Chloro-2-methoxy-N-pyridin-3-yl-benzenesulfonamide

Step 1: To chlorosulfuric acid (20 mL) was added1-bromo-2-chloro-4-methoxy-benzene (5.0 g, 23 mmol) dropwise at 0° C.The mixture was slowly warmed up to room temperature and was stirred atroom temperature for 2 h. TLC indicated the reaction was complete. Thenthe mixture was poured into ice-water. The aqueous layer was extractedwith EtOAc (20 mL×3). The extracts were dried over Na₂SO₄ and thesolution was filtered through a pad of silica gel, dried in vacuum toafford 2.0 g (yield: 27%) of 5-bromo-4-chloro-2-methoxy-benzenesulfonylchloride as white solid. ¹H NMR (DMSO-d6): δ=7.90 (1H, s), 7.24 (1H, s),3.79 (3H, s).

Step 2: The procedure is similar to Example I-1.

¹H NMR (DMSO-d6): δ=11.0 (1H, brs), 8.47-8.42 (2H, m), 8.04 (1H, t),7.90-7.80 (1H, m), 7.58-7.56 (1H, m), 7.56 (1H, s), 3.84 (3H, s). MS:m/z 378.8 (M+H⁺).

Step 3: The mixture of5-bromo-4-chloro-2-methoxy-N-pyridin-3-yl-benzenesulfonamide (200 mg,0.53 mmol), 10% Pd/C (40 mg) in EtOH/DMF (10 mL/10 mL) was stirred atroom temperature under hydrogen balloon pressure for 2 days. The mixturewas filtered and the filtrate was concentrated in vacuum to afford crudecompound, which was purified by prep-HPLC to afford 40 mg of (25% yield)4-chloro-2-methoxy-N-pyridin-3-yl-benzenesulfonamide as white solid.

¹H NMR (DMSO-d6): δ=10.62 (1H, brs), 8.35 (1H, s), 8.28 (1H, d), 7.79(1H, d), 7.60 (1H, d), 7.41 (1H, t), 7.31 (1H, d), 7.13 (1H, dd), 3.88(3H, s). MS: m/z 398.9 (M+H⁺).

Example I-12 3-Chloro-4-methoxy-N-pyridin-3-yl-benzenesulfonamide

Step 1: The procedure is similar to step 1, Example I-10. ¹H NMR(DMSO-d6): δ=8.01 (1H, s), 7.52 (1H, d), 6.98 (1H, d).

Step 2: The procedure is similar to Example I-1.

¹H NMR (DMSO-d6): δ=10.74 (1H, brs), 8.33-8.30 (2H, m), 7.79 (1H, d),7.71 (1H, dd), 7.60-7.56 (1H, m), 7.38 (1H, dd), 7.31 (1H, d), 3.92 (3H,s). MS: m/z 299.0 (M+H⁺).

Example I-13 3-Chloro-2-methoxy-N-pyridin-3-yl-benzenesulfonamide

Step 1: To the mixture of 1-chloro-2-methoxy-benzene (1.75 g, 12.3 mmol)in THF (40 mL), was added s-BuLi (11.3 mL, 14.7 mmol) dropwise at −78°C. The mixture was stirred at −78° C. for 1 h. And then, SO₂ wasinjected with a balloon. The mixture was slowly warmed up to roomtemperature and stirred overnight. The reaction was diluted withanhydrous DCM (10 mL). NCS (4.9 g, 37 mmol) was added to the mixtureportion-wise at 0° C. and the reaction was slowly warmed up to roomtemperature. The solvent was evaporated in vacuum. The residue waspurified by silica gel column to afford 800 mg (yield: 27%) of3-chloro-2-methoxy-benzenesulfonyl chloride. ¹H NMR (DMSO-d6):δ=7.84-7.81 (2H, m), 7.10 (1H, d), 3.87 (3H, s).

Step 2: The procedure is similar to Example I-1.

¹H NMR (DMSO-d6): δ=10.90 (1H, brs), 8.32 (1H, d), 8.23 (1H, d), 7.65(1H, dd), 7.44-7.40 (3H, m), 7.27 (1H, dd), 3.90 (3H, s). MS: m/z 349.0(M+H⁺).

Example I-14 4-Methoxy-3-(pyridin-3-ylsulfamoyl)-benzoic acid methylester

Step 1: To chlorosulfuric acid (40 mL) was added 4-methoxy-benzoic acidmethyl ester (20 g, 0.12 mol) dropwise at 0° C. The mixture was thenslowly warmed up to room temperature and was stirred at room temperatureovernight. TLC indicated the reaction was complete. The mixture waspoured into ice-water and extracted with EtOAc (200 mL×3). The extractswere dried over Na₂SO₄ and concentrated in vacuum to afford crude methyl3-(chlorosulfonyl)-4-methoxybenzoate, which was purified by silica-gelchromatography (PE/EA, 100/1 to 10/1) to afford 1.6 g (5% yield) ofmethyl 3-(chlorosulfonyl)-4-methoxybenzoate as white solid. ¹H NMR(CDCl₃): δ=8.65 (1H, s), 8.39 (1H, d), 7.18 (1H, d), 4.17 (3H, s), 3.94(3H, s).

Step 2: This compound was prepared as described in Example I-1.

¹H NMR (DMSO-d6): δ=10.47 (1H, brs), 8.29 (1H, d), 8.21 (1H, dd),8.13-8.08 (2H, m), 7.47 (1H, d), 7.32 (1H, dd), 7.25-7.22 (1H, m), 3.95(3H, s), 3.83 (3H, s). MS: m/z 323.0 (M+H⁺).

Example I-15 4-Methoxy-3-(pyridin-3-ylsulfamoyl)-benzamide

Methyl 3-(chlorosulfonyl)-4-methoxybenzoate (100 mg, 0.31 mmol) andaqueous ammonia (2 mL) was heated in a sealed vessel at 120° C. for 18hours. The reaction mixture was cooled to room temperature andconcentrated to dryness. The residue was purified by prep-HPLC to afford25 mg (yield: 26%) of 4-methoxy-3-(pyridin-3-ylsulfamoyl)-benzamide aswhite solid.

¹H NMR (DMSO-d6): δ=10.39 (1H, brs), 8.31 (2H, s), 8.19 (1H, d), 8.07(2H, dd), 7.47 (1H, d), 7.42 (1H, s), 7.29-7.22 (2H, m), 3.92 (3H, s).MS: m/z 307.9 (M+H⁺).

Example I-16 4-Methoxy-N-methyl-3-(pyridin-3-ylsulfamoyl)-benzamide

Methyl 3-(chlorosulfonyl)-4-methoxybenzoate (100 mg, 0.31 mmol) andMeNH₂ alcohol solution (3 mL) was heated in a sealed vessel at 120° C.for 18 hours. The reaction mixture was cooled to room temperature andconcentrated to dryness. The residue was purified by prep-HPLC to afford32 mg (yield: 33%) of 4-methoxy-3-(pyridin-3-ylsulfamoyl)-benzamide aswhite solid.

¹H NMR (DMSO-d6): δ=10.60 (1H, brs), 8.58 (1H, d), 8.31-8.26 (3H, m),8.07 (1H, d), 7.63 (1H, d), 7.49 (1H, dd), 7.26 (1H, d), 3.92 (3H, s),2.76 (3H, d). MS: m/z 322.0 (M+H⁺).

Example I-17 N-Ethyl-4-methoxy-3-(pyridin-3-ylsulfamoyl)-benzamide

Step 1: The mixture of 4-methoxy-3-(pyridin-3-ylsulfamoyl)-benzoic acidmethyl ester (1.5 g, 4.65 mmol), LiOH (0.45 g, 18.6 mmol) in THF/H₂O (10mL/10 mL) was stirred at 50° C. for 2 h. THF was evaporated in vacuum.The aqueous layer was extracted with EtOAc (30 mL×3). The extracts weredried over Na₂SO₄ and concentrated in vacuum to afford 1.1 g of (yield:77%) 4-methoxy-3-(N-(pyridin-3-yl)sulfamoyl)benzoic acid as yellowsolid.

Step 2: The mixture of 4-methoxy-3-(N-(pyridin-3-yl)sulfamoyl)benzoicacid (100 mg, 0.32 mmol), HATU (127 mg, 0.34 mmol), DIPEA (124 mg, 0.96mmol) and ethyl amine HCl salt (52 mg, 0.64 mmol) in DCM (3 mL) wasstirred at room temperature overnight. The solvent was evaporated invacuum. The residue was purified by prep-HPLC to afford 32 mg (yield:30%) of N-ethyl-4-methoxy-3-(pyridin-3-ylsulfamoyl)-benzamide as yellowsolid.

¹H NMR (DMSO-d6): δ=10.39 (1H, brs), 9.61 (1H, t), 8.30-8.26 (2H, m),8.20 (1H, d), 8.07 (1H, dd), 7.48 (1H, d), 7.26-7.22 (2H, m), 3.92 (3H,s), 1.50 (2H, q), 1.10 (3H, t). MS: m/z 336.1 (M+H⁺).

Example I-18 4-Methoxy-N-propyl-3-(pyridin-3-ylsulfamoyl)-benzamide

This compound was prepared as described in Example 1-17.

¹H NMR (DMSO-d6): δ=10.39 (1H, brs), 9.61 (1H, t), 8.30-8.26 (2H, m),8.20 (1H, d), 8.07 (1H, dd), 7.48 (1H, d), 7.26-8.22 (2H, m), 3.94 (3H,s), 3.25-3.21 (2H, m), 1.53-1.50 (2H, m), 1.10 (3H, t). MS: m/z 350.1(M+H⁺).

Example I-19 4-Methoxy-N-phenyl-3-(pyridin-3-ylsulfamoyl)-benzamide

This compound was prepared as described in Example I-17.

¹H NMR (DMSO-d6): δ=10.45 (1H, brs), 10.35 (1H, s), 8.42 (1H, d), 8.34(1H, d), 8.22 (2H, d), 7.75 (2H, d), 7.52 (1H, dd), 7.37-7.34 (3H, m),7.26 (1H, t), 7.12 (1H, t), 3.97 (3H, s). MS: m/z 384.1 (M+H⁺).

Example I-20 N-Cyclohexyl-4-methoxy-3-(pyridin-3-ylsulfamoyl)-benzamide

This compound was prepared as described in Example I-17.

¹H NMR (DMSO-d6): δ=10.33 (1H, brs), 8.32 (1H, d), 8.29-8.25 (2H, m),8.16 (1H, d), 8.05 (1H, dd), 7.45 (1H, d), 7.23-7.19 (2H, m), 3.88 (3H,s), 3.31-3.27 (1H, m), 1.75-1.25 (10H, m). MS: m/z 390.1 (M+H⁺).

Example I-214-Methoxy-N-(2-methoxy-ethyl)-3-(pyridin-3-ylsulfamoyl)-benzamide

This compound was prepared as described in Example 1-17.

¹H NMR (DMSO-d6): δ=10.40 (1H, brs), 8.69 (1H, t), 8.34-8.30 (2H, m),8.20 (1H, dd), 8.00 (1H, dd), 7.50 (1H, d), 7.29-7.25 (2H, m), 3.95 (3H,s), 3.45-3.39 (4H, m), 3.35 (3H, s). MS: m/z 366.1 (M+H⁺).

Example I-222-Methoxy-5-(4-methyl-piperazine-1-carbonyl)-N-pyridin-3-yl-benzenesulfonamide

This compound was prepared as described in Example 1-17.

¹H NMR (DMSO-d6): δ=10.43 (1H, brs), 8.29 (1H, d), 8.21 (1H, dd),8.13-8.10 (1H, m), 7.47 (1H, d), 7.32 (1H, dd), 7.25-7.21 (2H, m), 3.92(3H, s), 3.57-3.17 (4H, m), 2.34-2.29 (4H, m), 2.20 (3H, s). MS: m/z391.1 (M+H⁺).

Example I-232-Methoxy-5-(morpholine-4-carbonyl)-N-pyridin-3-yl-benzenesulfonamide

This compound was prepared as described in Example I-17.

¹H NMR (DMSO-d6): δ=10.40 (1H, brs), 8.29 (1H, d), 8.22 (1H, dd),7.77-7.74 (1H, m), 7.64 (1H, dd), 7.48 (1H, d), 7.28-7.24 (2H, m), 3.91(3H, s), 3.57-3.37 (8H, m). MS: m/z 378.1 (M+H⁺).

Example II Example II-15-Chloro-2-methoxy-N-quinolin-3-yl-benzenesulfonamide

The mixture of 5-chloro-2-methoxy-benzenesulfonyl chloride (167 mg, 0.69mmol), quinolin-3-ylamine (100 mg, 0.69 mmol), DMAP (10 mg, cat.) inpyridine (5 mL) was stirred at 50° C. for 2 h. LCMS indicated thereaction was complete. The solvent was evaporated in vacuum. The residuewas triturated with DCM (5 mL). The suspension was collected byfiltration to give crude product, which was purified by prep-HPLC toafford 100 mg (42% yield) of5-chloro-2-methoxy-N-quinolin-3-yl-benzenesulfonamide as pale yellowsolid.

¹H NMR (CDCl₃): δ=8.54 (1H, brs), 8.03 (1H, s), 7.97 (1H, d), 7.80-7.77(2H, m), 7.65 (1H, t), 7.58 (1H, t), 7.41 (1H, d), 7.26 (1H, s), 6.96(1H, d), 4.05 (3H, s). MS: m/z 295.0 (M+H⁺).

Example II-22-Methoxy-N-quinolin-3-yl-5-trifluoromethyl-benzenesulfonamide

¹H NMR (DMSO-d6): δ=10.63 (1H, brs), 8.70 (1H, d), 8.08 (1H, d),7.91-7.85 (4H, m), 7.64 (1H, t), 7.55 (1H, t), 7.40 (1H, dd), 3.93 (3H,s). MS: m/z 382.9 (M+H⁺).

Example II-3 2-Methoxy-4-methyl-N-quinolin-3-yl-benzenesulfonamide

¹H NMR (CDCl₃): δ=8.52 (1H, brs), 8.03 (1H, s), 7.97 (1H, d), 7.75 (1H,d), 7.68 (1H, d), 7.62 (1H, t), 7.52 (1H, t), 7.29-7.25 (1H, m), 6.78(1H, s), 6.74 (1H, d), 4.04 (3H, s), 2.32 (1H, s). MS: m/z 329.0 (M+H⁺).

Example II-4 2,4-Dimethoxy-N-quinolin-3-yl-benzenesulfonamide

¹H NMR (DMSO-d6): δ=10.45 (1H, brs), 8.66 (1H, d), 7.89-7.84 (3H, m),7.75 (1H, d), 7.61 (1H, t), 7.53 (1H, t), 6.62 (1H, d), 6.55 (1H, dd),3.85 (3H, s), 3.75 (1H, s). MS: m/z 345.0 (M+H⁺).

Example II-5 5-Cyano-2-methoxy-N-quinolin-3-yl-benzenesulfonamide

This compound was prepared as described in Example I-6.

¹H NMR (DMSO-d6): δ=10.86 (1H, brs), 8.70 (1H, s), 8.25 (1H, s), 8.05(1H, d), 7.97-7.94 (3H, m), 7.62 (1H, t), 7.55 (1H, t), 7.36 (1H, dd),3.95 (3H, s). MS: m/z 340.0 (M+H⁺).

Example II-6 4-Methoxy-biphenyl-3-sulfonic acid quinolin-3-ylamide

This compound was prepared as described in Example I-7.

¹H NMR (DMSO-d6): δ=10.67 (1H, brs), 8.70 (1H, d), 8.05 (1H, d), 7.98(1H, d), 7.90-7.84 (3H, m), 7.60-7.53 (3H, m), 7.48 (1H, t), 7.46 (2H,t), 7.37-7.33 (1H, m), 7.25 (1H, dd) 3.89 (3H, s). MS: m/z 391.0 (M+H⁺).

Example II-7 5-Furan-3-yl-2-methoxy-N-quinolin-3-yl-benzenesulfonamide

¹H NMR (DMSO-d6): δ=10.64 (1H, brs), 8.70 (1H, d), 8.19 (1H, s),7.98-7.94 (2H, m), 7.91-7.87 (2H, m), 7.76 (1H, dd), 7.73 (1H, s), 7.62(1H, d), 7.52 (1H, d), 7.19 (1H, d), 6.93 (1H, s), 3.87 (3H, s). MS: m/z381.0 (M+H⁺).

Example II-82-Methoxy-N-quinolin-3-yl-5-thiophen-3-yl-benzenesulfonamide

¹H NMR (DMSO-d6): δ=10.67 (1H, brs), 8.70 (1H, d), 8.09 (1H, s),8.02-7.98 (2H, m), 7.86-7.81 (4H, m), 7.63-7.58 (2H, m), 7.50 (1H, d),7.20 (1H, d), 3.89 (3H, s). MS: m/z 397.0 (M+H⁺).

Example II-9 2-Methoxy-5-pyridin-4-yl-N-quinolin-3-yl-benzenesulfonamide

This compound was prepared as described in Example I-10.

¹H NMR (DMSO-d6): δ=10.74 (1H, brs), 8.72 (1H, d), 8.62 (2H, d), 8.21(1H, d), 8.04-8.00 (2H, m), 7.90-7.86 (2H, m), 7.71-7.68 (2H, m), 7.61(1H, t), 7.51 (1H, t), 7.31 (1H, t), 3.93 (3H, s). MS: m/z 392.0 (M+H⁺).

Example II-10 4-Chloro-2-methoxy-N-quinolin-3-yl-benzenesulfonamide

Step 1: The procedure tobromo-4-chloro-2-methoxy-N-quinolin-3-yl-benzenesulfonamide is similarto Example II-1.

¹H NMR (DMSO-d6): δ=10.82 (1H, brs), 8.88 (1H, d), 8.04 (1H, s), 7.98(1H, s), 7.92-7.88 (2H, m), 7.83 (1H, t), 7.56 (1H, t), 7.51 (1H, s),3.88 (3H, s). MS: m/z 428.8 (M+H⁺).

Step 2: To the mixture of5-bromo-4-chloro-2-methoxy-N-quinolin-3-yl-benzenesulfonamide (100 mg,0.23 mmol) in THF (3 mL) was added BuLi (0.3 mL, 2.5M in THF) dropwiseat −78° C., and the mixture was stirred for another 3 h. The mixture wasquenched with water and concentrated to give a crude product, which waspurified by prep-HPLC to afford 40 mg (yield: 50%) of4-chloro-2-methoxy-N-quinolin-3-yl-benzenesulfonamide as white solid.

¹H NMR (DMSO-d6): δ=10.69 (1H, brs), 8.88 (1H, s), 8.04-7.99 (3H, m),7.81 (1H, d), 7.63 (1H, t), 7.55 (1H, t), 7.29 (1H, s), 7.10 (1H, d),3.89 (3H, s). MS: m/z 349.0 (M+H⁺).

Example II-11 3-Chloro-4-methoxy-N-quinolin-3-yl-benzenesulfonamide

This compound was prepared as described in Example I-12.

¹H NMR (DMSO-d6): δ=10.74 (1H, brs), 8.62 (1H, d), 8.02 (1H, d),7.97-7.94 (2H, m), 7.86 (1H, d), 7.73 (1H, dd), 7.65 (1H, t), 7.60 (1H,t), 7.26 (1H, d), 3.93 (3H, s). MS: m/z 349.0 (M+H⁺).

Example II-12 3-Chloro-2-methoxy-N-quinolin-3-yl-benzenesulfonamide

This compound was prepared as described in Example I-13.

¹H NMR (DMSO-d6): δ=11.22 (1H, brs), 8.77 (1H, d), 7.97-7.93 (3H, m),7.77 (1H, d), 7.70 (1H, t), 7.62-7.59 (1H, m), 7.51 (1H, t), 7.45 (1H,t), 3.91 (3H, s). MS: m/z 349.0 (M+H⁺).

Example II-13 Methyl 4-methoxy-3-(N-(quinolin-3-yl)sulfamoyl)benzoate

This compound was prepared as described in Example I-14.

¹H NMR (DMSO-d6): δ=10.74 (1H, brs), 8.67 (1H, d), 8.33 (1H, d), 8.11(1H, dd), 7.95 (1H, d), 7.88 (2H, d), 7.63 (1H, m), 7.50-7.46 (1H, m),7.31-7.28 (1H, m), 3.95 (3H, s), 3.81 (3H, s). MS: m/z 373.0 (M+H⁺).

Example II-14 4-Methoxy-3-(quinolin-3-ylsulfamoyl)-benzamide

This compound was prepared as described in Example I-15.

¹H NMR (DMSO-d6): δ=10.69 (1H, brs), 8.69 (1H, d), 8.37 (1H, d),8.05-8.01 (2H, m), 7.96 (1H, d), 7.91-7.87 (2H, m), 7.63-7.60 (1H, m),7.50-7.47 (1H, m), 7.29-7.26 (1H, m), 7.23 (1H, d), 3.9 (3H, s). MS: m/z358.0 (M+H⁺).

Example II-15 4-Methoxy-N-methyl-3-(quinolin-3-ylsulfamoyl)-benzamide

This compound was prepared as described in Example I-16.

¹H NMR (DMSO-d6): δ=8.44-8.35 (3H, m), 7.84 (1H, dd), 7.71 (1H, t),7.55-7.50 (2H, m), 7.33-7.29 (2H, m), 7.06 (1H, d), 7.00 (1H, brs), 3.76(3H, s), 2.36 (3H, s). MS: m/z 372.0 (M+H⁺).

Example II-16 N-Ethyl-4-methoxy-3-(quinolin-3-ylsulfamoyl)-benzamide

Step 1: The mixture of 4-methoxy-3-(quinolin-3-ylsulfamoyl)-benzoic acidmethyl ester (2.6 g, 7.0 mmol), LiOH (1.5 g, 35 mmol) in THF/H₂O (10mL/10 mL) was stirred at 50° C. for 2 h. LCMS indicated the reaction wascomplete. THF was evaporated in vacuum. The aqueous layer was extractedwith EtOAc (30 mL×3). The extracts were dried over Na₂SO₄ andconcentrated in vacuum to afford 1.8 g of (yield: 72%)4-methoxy-3-(quinolin-3-ylsulfamoyl)-benzoic acid as yellow solid. MS:m/z 357.1 (M−H⁺).

Step 2: The mixture of 4-methoxy-3-(quinolin-3-ylsulfamoyl)-benzoic acid(100 mg, 0.28 mmol), HATU (127 mg, 0.34 mmol), DIPEA (72 mg, 0.56 mmol)and ethyl amine HCl salt (46 mg, 0.56 mmol) in DCM (3 mL) was stirred atroom temperature overnight. The solvent was evaporated in vacuum. Theresidue was purified by prep-HPLC to afford 30 mg (yield: 28%) ofN-ethyl-4-methoxy-3-(quinolin-3-ylsulfamoyl)-benzamide as yellow solid.¹H NMR (DMSO-d6): δ=10.87 (1H, brs), 8.67 (1H, d), 8.56 (1H, t), 8.33(1H, d), 8.04 (1H, dd), 7.95 (1H, d), 7.88-7.84 (2H, m), 7.61-7.58 (1H,m), 7.54-7.51 (1H, m), 7.25 (1H, d), 3.91 (3H, s), 3.24 (2H, q), 1.11(3H, t). MS: m/z 386.1 (M+H⁺).

Example II-17 4-Methoxy-N-propyl-3-(quinolin-3-ylsulfamoyl)-benzamide

This compound was prepared as described in Example II-16.

¹H NMR (DMSO-d6): δ=10.68 (1H, brs), 8.69 (1H, d), 8.60-8.55 (1H, m),8.35 (1H, d), 8.03 (1H, dd), 7.95 (1H, d), 7.88-7.84 (2H, m), 7.63 (1H,t), 7.54 (1H, t), 7.24 (1H, d), 3.91 (3H, s), 3.16 (2H, q), 1.50-1.46(2H, m), 0.86 (3H, t). MS: m/z 400.1 (M+H⁺).

Example II-18 4-Methoxy-N-phenyl-3-(quinolin-3-ylsulfamoyl)-benzamide

This compound was prepared as described in Example II-16.

¹H NMR (DMSO-d6): δ=10.72 (1H, brs), 10.32 (1H, brs), 8.71 (1H, d), 8.46(1H, d), 8.20 (1H, dd), 7.97-7.94 (1H, m), 7.88-7.84 (2H, m), 7.70-7.66(2H, m), 7.62 (1H, m), 7.55-7.52 (1H, m), 7.32-7.28 (3H, m), 7.10-7.07(1H, m), 3.95 (3H, s). MS: m/z 434.1 (M+H⁺).

Example II-19N-Cyclohexyl-4-methoxy-3-(quinolin-3-ylsulfamoyl)-benzamide

This compound was prepared as described in Example II-16.

¹H NMR (DMSO-d6): δ=10.66 (1H, brs), 8.69 (1H, d), 8.34-8.30 (2H, m),8.04 (1H, d), 7.95 (1H, d), 7.86-7.82 (2H, m), 7.63 (1H, t), 7.52 (1H,t), 7.21 (1H, d), 3.91 (3H, s), 3.72-3.68 (1H, m), 1.72-1.62 (4H, m),1.57-1.53 (1H, m), 1.20-1.16 (4H, m), 1.10-1.07 (1H, m). MS: m/z 440.1(M+H⁺).

Example II-204-Methoxy-N-(2-methoxy-ethyl)-3-(quinolin-3-ylsulfamoyl)-benzamide

This compound was prepared as described in Example II-16.

¹H NMR (DMSO-d6): δ=10.89 (1H, brs), 8.70-8.66 (2H, m), 8.36 (1H, d),8.08 (1H, d), 7.97-7.93 (1H, m), 7.88-7.84 (2H, m), 7.65-7.62 (1H, m),7.54-7.51 (1H, m), 7.26 (1H, d), 3.95 (3H, s), 3.40-3.32 (4H, m), 3.20(3H, s). MS: m/z 416.1 (M+H⁺).

Example II-21N-(2-Dimethylamino-ethyl)-4-methoxy-3-(quinolin-3-ylsulfamoyl)-benzamide

This compound was prepared as described in Example II-16.

¹H NMR (CD₃OD): δ=8.70 (1H, s), 8.43 (1H, d), 8.09-8.04 (2H, m), 7.93(1H, d), 7.84 (1H, d), 7.70 (1H, t), 7.60 (1H, t), 7.30 (1H, d), 4.08(3H, s), 3.72 (2H, t), 3.37 (2H, t), 2.96 (6H, s). MS: m/z 429.1 (M+H⁺).

Example II-222-Methoxy-5-(4-methyl-piperazine-1-carbonyl)-N-quinolin-3-yl-benzenesulfonamide

This compound was prepared as described in Example II-16.

¹H NMR (CD₃OD): δ=8.77 (1H, d), 8.20 (1H, d), 8.05 (1H, d), 7.98 (1H,d), 7.91 (1H, d), 7.76-7.73 (1H, m), 7.65-7.62 (2H, m), 7.26 (1H, d),4.00 (3H, s), 3.60-3.30 (8H, m), 2.94 (3H, s). MS: m/z 441.1 (M+H⁺).

Example II-232-Methoxy-5-(morpholine-4-carbonyl)-N-quinolin-3-yl-benzenesulfonamide

This compound was prepared as described in Example II-16.

¹H NMR (CD₃OD): δ=8.67 (1H, d), 8.13 (1H, d), 7.90-7.86 (3H, m), 7.70(1H, t), 7.60 (2H, m), 7.22 (1H, d), 3.98 (3H, s), 3.50-2.50 (8H, m).MS: m/z 428.1 (M+H⁺).

Example III Example III-15-Bromo-2-methoxy-N-[5-(4-methoxy-phenyl)-pyridin-3-yl]-benzenesulfonamide

Step 1: A mixture of 5-bromopyridin-3-amine (300 mg, 1.74 mmol),4-methoxyphenylboronic acid (395 mg, 2.60 mmol), K₂CO₃ (240 mg, 5.10mmol) and Pd(PPh₃)₄ (197 mg, 0.17 mmol) in DMF/H₂O (5 ml/1 ml) waspurged with N₂ for 20 min. Then the mixture was stirred at 120° C. undermicrowave irridation for 10 min. After cooled to room temperature, thesolvent was removed in vacuum. The residue was diluted with EtOAc (30mL). The mixture was washed with water, brine and dried over Na₂SO₄. Thesolution was evaporated to dryness and purified by silica gel column(DCM/MeOH, 1/0-40/1) to afford 264 mg (yield: 44%) of5-(4-methoxyphenyl)pyridin-3-amine as white solid. MS: m/z 201.1 (M+H⁺).

Step 2: A mixture of 5-(4-methoxyphenyl)pyridin-3-amine (70 mg, 0.35mmol), 2,5-dimethoxybenzene-1-sulfonyl chloride (83 mg, 0.35 mmol) andDMAP (51 mg, 0.42 mmol) in pyridine (2 ml) was heated at 90° C. for 18h. After cooled to room temperature, the solvent was removed in vacuum.The residue was diluted with EtOAc (20 mL). The mixture was washed withwater, brine and dried over Na₂SO₄. The solution was evaporated todryness and the residue was purified by prep-HPLC to afford 25 mg(yield: 18%) of5-bromo-2-methoxy-N-[5-(4-methoxy-phenyl)-pyridin-3-yl]-benzenesulfonamideas off-white solid.

¹H NMR (DMSO-d6, 400 MHz): δ=10.42 (1H, brs), 8.48 (1H, s), 8.25 (1H,d), 7.65 (1H, s), 7.50 (2H, d), 7.32 (1H, d), 7.17-7.14 (2H, m), 7.05(2H, d), 3.81 (3H, s), 3.80 (3H, s), 3.72 (3H, s). MS: m/z 401.1 (M+H⁺).

Example III-25-Bromo-2-methoxy-N-[5-(4-methoxy-phenyl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.65 (1H, brs), 8.56 (1H, s), 8.29 (1H,d), 7.94 (1H, d), 8.35 (1H, dd), 7.69 (1H, s), 7.56 (2H, d), 7.22 (1H,d), 7.10 (2H, d), 3.91 (3H, s), 3.85 (3H, s). MS: m/z 448.9 (M+H⁺).

Example III-35-Chloro-2-methoxy-N-[5-(4-methoxy-phenyl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.58 (1H, brs), 8.50 (1H, s), 8.24 (1H,s), 7.78 (1H, s), 7.66-7.54 (2H, m), 7.51 (2H, d), 7.23 (1H, d), 7.06(2H, d), 3.86 (3H, s), 3.80 (3H, s). MS: m/z 404.9 (M+H⁺).

Example III-4N-(5-(4-(Benzyloxy)phenyl)pyridin-3-yl)-2,5-dimethoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO, 400 HMz): δ=10.41 (1H, brs), 8.48 (1H, d), 8.24 (1H, d),7.64 (1H, t), 7.51-7.31 (8H, m), 7.17-7.10 (4H, m), 5.16 (2H, s), 3.80(3H, s), 3.72 (3H, s). MS: m/z 477.1 (M+H⁺).

Example III-5N-(5-(4-(Benzyloxy)phenyl)pyridin-3-yl)-5-bromo-2-methoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.58 (1H, brs), 8.52 (1H, d), 8.23 (1H,d), 7.88 (1H, d), 7.80-7.76 (1H, m), 7.64 (1H, d), 7.51 (2H, d), 7.47(2H, d), 7.41 (2H, t), 7.35 (1H, d), 7.16 (1H, d), 7.13 (2H, d), 5.17(2H, s) 3.85 (3H, s). MS: m/z 525.0 (M+H⁺).

Example III-6N-(5-(4-(benzyloxy)phenyl)pyridin-3-yl)-5-chloro-2-methoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 HMz): δ=10.56 (1H, brs), 8.51 (1H, d), 8.24 (1H,d), 7.77 (1H, d), 7.64-7.53 (2H, m), 7.53-7.34 (7H, m), 7.24 (1H, d),7.13 (2H, d), 5.17 (2H, s), 3.86 (3H, s). MS: m/z 481.1 (M+H⁺).

Example III-7N-[5-(4-Hydroxy-phenyl)-pyridin-3-yl]-2,5-dimethoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 HMz): δ=9.70 (1H, brs), 8.37 (1H, d), 8.17 (1H, d),7.57 (1H, t), 7.37 (2H, d), 7.31 (1H, d), 7.13-7.09 (2H, m), 6.86 (2H,d), 3.78 (3H, s), 3.72 (3H, s). MS: m/z 387.0 (M+H⁺).

Example III-85-Bromo-N-[5-(4-hydroxy-phenyl)-pyridin-3-yl]-2-methoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 HMz): δ=9.71 (1H, brs), 8.42 (1H, s), 8.16 (1H, d),7.85 (1H, d), 7.74 (1H, dd), 7.55 (1H, t), 7.37-7.33 (2H, m), 7.15 (1H,d), 6.84 (2H, d), 3.82 (3H, s). MS: m/z 434.8 (M+H⁺).

Example III-95-Chloro-2-methoxy-N-(5-pyrimidin-2-yl-pyridin-3-yl)-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO, 400 HMz): δ=9.70 (1H, brs), 8.37 (1H, d), 8.17 (1H, d),7.57 (1H, t), 7.37 (2H, d), 7.31 (1H, d), 7.13-7.09 (2H, m), 6.86 (2H,d), 3.78 (3H, s), 3.72 (3H, s). MS: m/z 391.0 (M+H⁺).

Example III-102,5-Dimethoxy-N-(5-p-tolyl-pyridin-3-yl)-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.44 (1H, brs), 8.49 (1H, s), 8.27 (1H,d), 7.66 (1H, d), 7.45 (2H, d), 7.33-7.28 (3H, m), 7.17-7.13 (2H, m),3.80 (3H, s), 3.72 (3H, s), 2.35 (3H, s). MS: m/z 385.0 (M+H⁺).

Example III-115-Bromo-2-methoxy-N-(5-p-tolyl-pyridin-3-yl)-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.70 (1H, brs), 8.57 (1H, s), 8.30 (1H,s), 7.90 (1H, d), 7.80 (1H, dd), 7.72 (1H, d), 7.47 (2H, d), 7.32 (2H,d), 7.24 (1H, d), 3.84 (3H, s), 2.35 (3H, s). MS: m/z 432.9 (M+H⁺).

Example III-125-Chloro-2-methoxy-N-(5-p-tolyl-pyridin-3-yl)-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.67 (1H, brs), 8.56 (1H, s), 8.29 (1H,s), 7.79 (1H, s), 7.72 (1H, s), 7.67 (1H, dd), 7.47 (2H, d), 7.31 (2H,d), 7.24 (1H, d), 3.85 (3H, s), 2.35 (3H, s). MS: m/z 388.9 (M+H⁺).

Example III-132,5-Dimethoxy-N-(5-(4-(trifluoromethyl)phenyl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.53 (1H, brs), 8.59 (1H, d), 8.36 (1H,d), 7.87 (2H, d), 7.82-7.75 (3H, m), 7.32 (1H, d), 7.18-7.13 (2H, m),3.79 (3H, s), 3.72 (3H, s). MS: m/z 439.0 (M+H⁺).

Example III-145-Bromo-2-methoxy-N-(5-(4-(trifluoromethyl)phenyl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.68 (1H, brs), 8.63 (1H, s), 8.36 (1H,s), 7.91-7.75 (7H, m), 7.18 (1H, d), 3.84 (3H, s). MS: m/z 486.8 (M+H⁺).

Example III-155-Chloro-2-methoxy-N-(5-(4-(trifluoromethyl)phenyl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.68 (1H, brs), 8.62 (1H, s), 8.36 (1H,d), 7.89-7.76 (6H, m), 7.67 (1H, dd), 7.24 (1H, d), 3.85 (3H, s). MS:m/z 442.9 (M+H⁺).

Example III-16N-[5-(4-Fluoro-phenyl)-pyridin-3-yl]-2,5-dimethoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.47 (1H, brs), 8.50 (1H, s), 8.29 (1H,s), 7.68 (1H, s), 7.61 (2H, dd), 7.37-7.30 (3H, m), 7.17-7.13 (2H, m),3.79 (3H, s), 3.72 (3H, s). MS: m/z 389.0 (M+H⁺).

Example III-175-Bromo-N-[5-(4-fluoro-phenyl)-pyridin-3-yl]-2-methoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.62 (1H, brs), 8.53 (1H, s), 8.28 (1H,d), 7.89 (1H, d), 7.78 (1H, dd), 7.68-7.60 (3H, m), 7.34 (2H, t), 7.18(1H, d), 3.84 (3H, s). MS: m/z 436.9 (M+H⁺).

Example III-185-Chloro-N-[5-(4-fluoro-phenyl)-pyridin-3-yl]-2-methoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.57 (1H, brs), 8.53 (1H, s), 8.29 (1H,s), 7.78 (1H, s), 7.68-7.60 (4H, m), 7.34 (2H, t), 7.23 (1H, d), 3.85(3H, s). MS: m/z 392.9 (M+H⁺).

Example III-19N-[5-(4-Chloro-phenyl)-pyridin-3-yl]-2,5-dimethoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.49 (1H, brs), 8.53 (1H, s), 8.31 (1H,s), 7.70 (1H, s), 7.63-7.52 (4H, m), 7.32 (1H, d), 7.15-7.13 (2H, m),3.79 (3H, s), 3.72 (3H, s). MS: m/z 404.9 (M+H⁺).

Example III-205-Bromo-N-[5-(4-chloro-phenyl)-pyridin-3-yl]-2-methoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.65 (1H, brs), 8.55 (1H, s), 8.30 (1H,s), 7.89 (1H, s), 7.78 (1H, dd), 7.69 (1H, s), 7.63-7.54 (3H, m), 7.17(2H, d), 3.84 (3H, s). MS: m/z 452.8 (M+H⁺).

Example III-215-Chloro-N-[5-(4-chloro-phenyl)-pyridin-3-yl]-2-methoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.61 (1H, brs), 8.56 (1H, s), 8.30 (1H,s), 7.78 (1H, s), 7.77-7.54 (6H, m), 7.23 (1H, d), 3.85 (3H, s). MS: m/z408.9 (M+H⁺).

Example III-22N-[5-(4-Bromo-phenyl)-pyridin-3-yl]-2,5-dimethoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO, 400 HMz): δ=10.46 (1H, s), 8.52 (1H, d), 8.32 (1H, d),7.71-7.69 (3H, m), 7.53 (2H, dd), 7.31 (1H, d), 7.19-7.12 (2H, m), 3.79(3H, s), 3.72 (3H, s). MS: m/z 448.7 (M+H⁺).

Example III-235-Bromo-N-[5-(4-bromo-phenyl)-pyridin-3-yl]-2-methoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

1H NMR (DMSO, 400 HMz): δ=10.61 (1H, s), 8.56 (1H, d), 8.31 (1H, d),7.88 (1H, d), 7.78 (1H, dd), 7.71-7.69 (3H, m), 7.54 (2H, dd), 7.18 (1H,d), 3.84 (3H, s). MS: m/z 496.7 (M+H⁺).

Example III-24N-[5-(4-Bromo-phenyl)-pyridin-3-yl]-5-chloro-2-methoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO, 400 HMz): δ=10.60 (1H, brs), 8.56 (1H, d), 8.31 (1H, d),7.78 (1H, d), 7.68-7.71 (4H, m), 7.54 (2H, d), 7.24 (1H, d), 3.85 (3H,s). MS: m/z 452.8 (M+H⁺).

Example III-25N-(5-Phenyl-pyridin-3-yl)-2,5-Dimethoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.48 (1H, brs), 8.52 (1H, s), 8.30 (1H,d), 7.70 (1H, s), 7.57-7.42 (6H, m), 7.32 (1H, d), 7.18-7.11 (1H, m),3.80 (3H, s), 3.72 (3H, s). MS: m/z 371.0 (M+H⁺).

Example III-265-Bromo-2-methoxy-N-(5-phenyl-pyridin-3-yl)-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.65 (1H, brs), 8.54 (1H, d), 8.29 (1H,d), 7.89 (1H, d), 7.79 (1H, dd), 7.69 (1H, t), 7.58-7.43 (5H, m), 7.18(1H, d), 3.85 (3H, s). MS: m/z 418.9 (M+H⁺).

Example III-275-Chloro-2-methoxy-N-(5-phenyl-pyridin-3-yl)-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.63 (1H, brs), 8.55 (1H, d), 8.30 (1H,d), 7.79 (1H, d), 7.71-7.65 (7H, m), 7.24 (1H, d), 3.86 (3H, s). MS: m/z374.9 (M+H⁺).

Example III-284-[5-(2,5-Dimethoxy-benzenesulfonylamino)-pyridin-3-yl]-benzoic acidmethyl ester

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.52 (1H, brs), 8.59 (1H, d), 8.34 (1H,d), 8.06 (2H, d), 7.78-7.71 (3H, d), 7.32 (1H, d), 7.20-7.11 (2H, m),3.88 (3H, s), 3.79 (3H, s), 3.72 (3H, s). MS: m/z 429.0 (M+H⁺).

Example III-294-[5-(5-Bromo-2-methoxy-benzenesulfonylamino)-pyridin-3-yl]-benzoic acidmethyl ester

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.71 (1H, brs), 8.62 (1H, s), 8.35 (1H,s), 8.07 (2H, d), 7.91 (1H, d), 7.81-7.72 (4H, m), 7.18 (1H, d), 3.89(3H, s), 3.84 (3H, s). MS: m/z 476.9 (M+H⁺).

Example III-304-[5-(5-Chloro-2-methoxy-benzenesulfonylamino)-pyridin-3-yl]-benzoicacid methyl ester

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.69 (1H, brs), 8.63 (1H, s), 8.35 (1H,s), 8.06 (2H, d), 7.81-7.72 (4H, m), 7.67 (1H, dd), 7.24 (1H, d), 3.89(3H, s), 3.85 (3H, s). MS: m/z 432.9 (M+H⁺).

Example III-31N-(5-(4-Cyanophenyl)pyridin-3-yl)-2,5-dimethoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.56 (1H, brs), 8.60 (1H, s), 8.36 (1H,d), 7.97 (2H, d), 7.80-7.76 (3H, m), 7.32 (1H, d), 7.20-7.11 (2H, m),3.79 (3H, s), 3.72 (3H, s). MS: m/z 396.0 (M+H⁺).

Example III-325-Bromo-N-(5-(4-cyanophenyl)pyridin-3-yl)-2-methoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.70 (1H, brs), 8.63 (1H, d), 8.35 (1H,d), 7.98 (2H, d), 7.89 (1H, d), 7.82-7.75 (4H, m), 7.18 (1H, d), 3.83(3H, s). MS: m/z 443.9 (M+H⁺).

Example III-335-Chloro-N-(5-(4-cyanophenyl)pyridin-3-yl)-2-methoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.68 (1H, brs), 8.64 (1H, s), 8.36 (1H,d), 7.97 (2H, d), 7.82-7.76 (4H, m), 7.67 (1H, dd), 7.24 (1H, d), 3.84(3H, s). MS: m/z 399.9 (M+H⁺).

Example III-342,5-Dimethoxy-N-[5-(2-methoxy-phenyl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example III-1.

1H NMR (DMSO, 400 HMz): δ=10.39 (1H, brs), 8.29 (1H, d), 8.26 (1H, d),7.61 (1H, t), 7.39 (1H, d), 7.29 (1H, d), 7.12-7.23 (4H, m), 7.04 (1H,t), 3.81 (3H, s), 3.72 (6H, d). MS: m/z 401.0 (M+H⁺).

Example III-355-Bromo-2-methoxy-N-[5-(2-methoxy-phenyl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO, 400 HMz): δ=10.54 (1H, brs), 8.31 (1H, s), 8.25 (1H, d),7.86 (1H, d), 7.80 (1H, dd), 7.61 (1H, s), 7.39 (1H, t), 7.22 (2H, t),7.14 (1H, d), 7.06 (1H, d), 3.87 (3H, s), 3.74 (3H, s). MS: m/z 448.9(M+H⁺).

Example III-365-Chloro-2-methoxy-N-[5-(2-methoxy-phenyl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO, 400 HMz): δ=10.54 (1H, brs), 8.32 (1H, d), 8.26 (1H, d),7.75 (1H, d), 7.69 (1H, dd), 7.62 (1H, t), 7.42-7.38 (1H, m), 7.38-7.23(2H, m), 7.13 (1H, d), 7.05 (1H, t), 3.87 (3H, s), 3.73 (3H, s). MS: m/z448.9 (M+H⁺).

Example III-372,5-Dimethoxy-N-(5-(o-tolyl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.45 (1H, brs), 8.31 (1H, d), 8.18 (1H,d), 7.41 (1H, d), 7.33-7.26 (4H, m), 7.20-7.10 (3H, m), 3.81 (3H, s),3.72 (3H, s), 2.04 (3H, s). MS: m/z 385.0 (M+H⁺).

Example III-385-Bromo-2-methoxy-N-(5-(o-tolyl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.61 (1H, brs), 8.30 (1H, d), 8.20 (1H,d), 7.85-7.77 (2H, m), 7.41 (1H, t), 7.34-7.24 (3H, m), 7.19 (1H, d),7.12 (1H, d), 3.87 (3H, s), 2.06 (3H, s). MS: m/z 432.9 (M+H⁺).

Example III-395-Chloro-2-methoxy-N-(5-o-tolyl-pyridin-3-yl)-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.61 (1H, brs), 8.30 (1H, s), 8.19 (1H,s), 7.74 (1H, d), 7.71-7.65 (1H, m), 7.40 (1H, s), 7.32-7.22 (4H, m),7.12 (1H, d), 3.87 (3H, s), 2.05 (3H, s). MS: m/z 389.0 (M+H⁺).

Example III-40N-[5-(2-Chloro-phenyl)-pyridin-3-yl]-2,5-dimethoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.50 (1H, brs), 8.35 (1H, d), 8.24 (1H,d), 7.61-7.55 (2H, m), 7.47-7.33 (2H, m), 7.37 (1H, t), 7.29 (1H, d),7.18-7.12 (2H, m), 3.80 (3H, s), 3.72 (3H, s). MS: m/z 404.9 (M+H⁺).

Example III-415-Bromo-N-[5-(2-chloro-phenyl)-pyridin-3-yl]-2-methoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.67 (1H, brs), 8.35 (1H, d), 8.27 (1H,d), 7.85 (1H, s), 7.79 (1H, dd), 7.61-7.36 (5H, m), 7.19 (1H, d), 3.87(3H, s). MS: m/z 452.8 (M+H⁺).

Example III-425-Chloro-N-[5-(2-chloro-phenyl)-pyridin-3-yl]-2-methoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.67 (1H, brs), 8.34 (1H, d), 8.25 (1H,d), 7.74 (1H, d), 7.70-7.37 (6H, m), 7.24 (1H, d), 3.87 (3H, s). MS: m/z408.9 (M+H⁺).

Example III-432,5-Dimethoxy-N-[5-(2-trifluoromethyl-phenyl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.50 (1H, brs), 8.35 (1H, d), 8.13 (1H,s), 7.85 (1H, d), 7.75 (1H, t), 7.68 (1H, t), 7.44 (1H, s), 7.35 (1H,d), 7.25 (1H, d), 7.17 (1H, d), 7.13 (1H, d), 3.80 (3H, s), 3.70 (3H,s). MS: m/z 439.0 (M+H⁺).

Example III-445-Bromo-2-methoxy-N-[5-(2-trifluoromethyl-phenyl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.66 (1H, brs), 8.35 (1H, d), 8.17 (1H,s), 7.88-7.66 (5H, m), 7.44 (1H, s), 7.37 (1H, d), 7.18 (1H, d), 3.86(3H, s). MS: m/z 486.9 (M+H⁺).

Example III-455-Bromo-2-methoxy-N-[5-(2-trifluoromethyl-phenyl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.66 (1H, brs), 8.37 (1H, s), 8.18 (1H,s), 7.85 (1H, d), 7.78-7.64 (4H, m), 7.45 (1H, s), 7.37 (1H, d), 7.24(1H, d), 3.870 (3H, s). MS: m/z 442.9 (M+H⁺).

Example III-462,5-Dimethoxy-N-[5-(3-methoxy-phenyl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 HMz): δ=10.47 (1H, brs), 8.52 (1H, d), 8.30 (1H,d), 7.67 (1H, d), 7.41 (1H, t), 7.33 (1H, d), 7.06-7.20 (4H, m), 6.99(1H, dd), 3.81 (6H, d), 3.72 (3H, s). MS: m/z 401.0 (M+H⁺).

Example III-475-Bromo-2-methoxy-N-[5-(3-methoxy-phenyl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 HMz): δ=10.61 (1H, brs), 8.55 (1H, d), 8.30 (1H,d), 7.88 (1H, d), 7.79 (1H, dd), 7.67 (1H, t), 7.41 (1H, d), 7.19 (1H,d), 7.09 (2H, t), 7.00 (1H, dd), 3.85 (3H, s), 3.82 (3H, s). MS: m/z448.9 (M+H⁺).

Example III-485-Chloro-2-methoxy-N-[5-(3-methoxy-phenyl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 HMz): δ=10.60 (1H, brs), 8.56 (1H, d), 8.30 (1H,d), 7.78 (1H, d), 7.66-7.69 (2H, m), 7.42 (1H, t), 7.25 (1H, d), 7.12(2H, t), 7.01 (1H, dd), 3.86 (3H, s), 3.82 (3H, s). MS: m/z 404.9(M+H⁺).

Example III-492,5-Dimethoxy-N-(5-m-tolyl-pyridin-3-yl)-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.47 (1H, brs), 8.47 (1H, d), 8.28 (1H,d), 7.66 (1H, d), 7.39-7.32 (4H, m), 7.27-7.13 (3H, m), 3.80 (3H, s),3.73 (3H, s), 2.37 (3H, s). MS: m/z 385.0 (M+H⁺).

Example III-505-Bromo-2-methoxy-N-(5-m-tolyl-pyridin-3-yl)-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.63 (1H, brs), 8.52 (1H, d), 8.29 (1H,d), 7.91 (1H, d), 7.79 (1H, dd), 7.66 (1H, t), 7.40-7.34 (3H, m),7.26-7.16 (2H, m), 3.85 (3H, s), 2.38 (3H, s). MS: m/z 432.9 (M+H⁺).

Example III-515-Chloro-2-methoxy-N-(5-m-tolyl-pyridin-3-yl)-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.62 (1H, brs), 8.52 (1H, d), 8.28 (1H,d), 7.80 (1H, d), 7.71-7.65 (2H, m), 7.39-7.34 (3H, m), 7.26-7.23 (2H,m), 3.86 (3H, s), 2.38 (3H, s). MS: m/z 389.0 (M+H⁺).

Example III-52N-[5-(3-Chloro-phenyl)-pyridin-3-yl]-2,5-dimethoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.53 (1H, brs), 8.56 (1H, s), 8.33 (1H,d), 7.72 (1H, s), 7.62 (1H, s), 7.53-7.51 (3H, m), 7.33 (1H, d),7.19-7.12 (2H, m), 3.79 (3H, s), 3.73 (3H, s). MS: m/z 405.0 (M+H⁺).

Example III-535-Bromo-N-[5-(3-chloro-phenyl)-pyridin-3-yl]-2-methoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.64 (1H, brs), 8.58 (1H, d), 8.32 (1H,d), 7.90 (1H, d), 7.79 (1H, dd), 7.70 (1H, t), 7.65 (1H, s), 7.56-7.50(3H, m), 7.18 (1H, d), 3.84 (3H, s). MS: m/z 452.8 (M+H⁺).

Example III-545-Chloro-N-[5-(3-chloro-phenyl)-pyridin-3-yl]-2-methoxy-benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.64 (1H, brs), 8.58 (1H, d), 8.32 (1H,d), 7.79 (1H, d), 7.72-7.63 (3H, m), 7.55-7.50 (3H, m), 7.24 (1H, d),3.85 (3H, s). MS: m/z 408.9 (M+H⁺).

Example III-55N-(5-(2,4-Dimethoxyphenyl)pyridin-3-yl)-2,5-dimethoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO, 400 HMz): δ=10.33 (1H, brs), 8.25 (1H, d), 8.21 (1H, d),7.58 (1H, d), 7.28 (1H, d), 7.14-7.18 (3H, m), 6.66 (1H, d), 6.62 (1H,dd), 3.81 (3H, s), 3.80 (3H, s), 3.72 (6H, d). MS: m/z 431.0 (M+H⁺).

Example III-565-Bromo-N-(5-(2,4-dimethoxyphenyl)pyridin-3-yl)-2-methoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.50 (1H, brs), 8.28 (1H, d), 8.20 (1H,d), 7.84 (1H, d), 7.79 (1H, dd), 7.58 (1H, s), 7.22-7.15 (2H, m), 6.67(1H, d), 6.65-6.60 (1H, m), 3.87 (3H, s), 3.81 (3H, s), 3.76 (3H, s).MS: m/z 478.9 (M+H⁺).

Example III-575-Chloro-N-(5-(2,4-dimethoxyphenyl)pyridin-3-yl)-2-methoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO, 400 HMz): δ=10.51 (1H, brs), 8.27 (1H, d), 8.21 (1H, d),7.74 (1H, d), 7.68 (1H, dd), 7.58 (1H, t), 7.26 (1H, d), 7.17 (1H, d),6.67 (1H, d), 6.63 (1H, dd), 3.87 (3H, s), 3.80 (3H, s), 3.73 (3H, s).MS: m/z 434.9 (M+H⁺).

Example III-58N-([2,3′-Bipyridin]-5′-yl)-2,5-dimethoxybenzenesulfonamide

Step 1, 2: 5-Bromo-pyridin-3-ylamine (1.0 g, 5.8 mmol),bis(pinacolato)diboron (1.46 g, 5.7 mmol), Pd(dppf)Cl₂ (200 mg), AcOK(1.1 g, 11.4 mmol) were stirred in 1,4-dioxane (15 mL) at 100° C. underN₂ for 4 h. After cooled to room temperature, to the mixture was added2-bromo-pyridine (0.91 g, 5.7 mmol), Cs₂CO₃ (7.4 g, 22.8 mmol),Pd(PPh₃)₄ (200 mg), water (3 mL). The mixture was then heated to 100° C.under N₂ for 2 h. The solvent was concentrated under reduced pressure.The residue was dissolved in water and the aqueous phase was extractedwith EtOAc (30 mL×3). The organic layer was washed with brine, driedover anhydrous Na₂SO₄ and concentrated to dryness under reducedpressure. The crude was purified via silic gel column (DCM/MeOH, 20/1)to afford 300 mg (2-step yield: 30%) of [2,3]Bipyridinyl-5′-ylamine. MS:m/z 172.0 (M+H⁺).

Step 3: This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d₆, 400 HMz): δ=10.48 (1H, brs), 8.87 (1H, s), 8.69 (1H,d), 8.36 (1H, s), 8.22 (1H, s), 7.91-7.95 (2H, m), 7.32-7.43 (2H, m),7.13-7.14 (2H, m), 3.80 (3H, s), 3.72 (3H, s). MS: m/z 372.1 (M+H⁺).

Example III-59N-([2,3′-Bipyridin]-5′-yl)-5-bromo-2-methoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d₆, 400 HMz): δ=10.63 (1H, brs), 8.91 (1H, s), 8.71 (1H,d), 8.37 (1H, s), 8.22 (1H, s), 7.98-7.75 (4H, m), 7.41-7.44 (1H, m),7.17 (1H, d), 3.85 (3H, s). MS: m/z 420.1 (M+H⁺).

Example III-60N-([2,3′-Bipyridin]-5′-yl)-5-chloro-2-methoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d₆, 400 HMz): δ=10.62 (1H, brs), 8.90 (1H, s), 8.70 (1H,d), 8.36 (1H, s), 8.21 (1H, s), 7.98-7.89 (2H, m), 7.76 (1H, d),7.66-7.64 (1H, m), 7.40-7.44 (1H, m), 7.22-7.24 (1H, m), 3.85 (3H, s).MS: m/z 376.0 (M+H⁺).

Example III-61 N-([3,3′-Bipyridin]-5-yl)-2,5-dimethoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.54 (1H, brs), 8.77 (1H, d), 8.63 (1H,dd), 8.58 (1H, d), 8.35 (1H, d), 8.00 (1H, dt), 7.74 (1H, t), 7.53 (1H,dd), 7.33 (1H, d), 7.20-7.12 (2H, m), 3.80 (3H, s), 3.72 (3H, s). MS:m/z 372.0 (M+H⁺).

Example III-62N-([3,3′-Bipyridin]-5-yl)-5-bromo-2-methoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.68 (1H, brs), 8.80 (1H, s), 8.64 (1H,d), 8.62 (1H, s), 8.35 (1H, d), 8.01 (1H, d), 7.90 (1H, d), 7.79 (1H,dd), 7.74 (1H, s), 7.56-7.50 (1H, m), 7.18 (1H, d), 3.85 (3H, s). MS:m/z 419.9 (M+H⁺).

Example III-63N-([3,3′-Bipyridin]-5-yl)-5-chloro-2-methoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.68 (1H, brs), 8.80 (1H, d), 8.62 (1H,dd), 8.60 (1H, dd), 8.34 (1H, d), 8.01 (1H, dt), 7.75 (2H, dd), 7.67(1H, dd), 7.55-7.50 (1H, m), 7.24 (1H, d), 3.85 (3H, s). MS: m/z 375.9(M+H⁺).

Example III-64 N-([3,4′-Bipyridin]-5-yl)-2,5-dimethoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.58 (1H, brs), 8.69-8.64 (3H, m), 8.38(1H, s), 7.81 (1H, s), 7.63-7.60 (2H, m), 7.33 (1H, s), 7.20-7.12 (2H,m), 3.79 (3H, s), 3.72 (3H, s). MS: m/z 372.0 (M+H⁺).

Example III-65N-([3,4′-Bipyridin]-5-yl)-5-bromo-2-methoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO, 400 HMz): δ=10.72 (1H, brs), 8.72-8.65 (3H, m), 8.38 (1H,d), 7.91 (1H, d), 7.81-7.77 (2H, m), 7.63 (2H, dd), 7.18 (1H, d), 3.84(3H, s). MS: m/z 419.9 (M+H⁺).

Example III-66N-([3,4′-Bipyridin]-5-yl)-5-chloro-2-methoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.76 (1H, brs), 8.82 (2H, d), 8.76 (1H,d), 8.42 (1H, d), 7.92-7.88 (3H, m), 7.80 (1H, d), 7.68 (1H, dd), 7.24(1H, d), 3.84 (3H, s). MS: m/z 375.9 (M+H⁺).

Example III-67N-(5-(Furan-2-yl)pyridin-3-yl)-2,5-dimethoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 HMz): δ=10.49 (1H, brs), 8.59 (1H, d), 8.20 (1H,d), 7.84 (1H, d), 7.74 (1H, t), 7.31 (1H, d), 7.16-7.04 (3H, m), 6.63(1H, dd), 3.80 (3H, s), 3.72 (3H, s). MS: m/z 360.9 (M+H⁺).

Example III-685-Bromo-N-(5-(furan-2-yl)pyridin-3-yl)-2-methoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 HMz): δ=10.63 (1H, brs), 8.63 (1H, d), 8.19 (1H,d), 7.72-7.87 (4H, m), 7.18 (1H, d), 7.17 (1H, d), 6.64 (1H, dd), 3.85(3H, s). MS: m/z 408.8 (M+H⁺).

Example III-695-Chloro-N-(5-(furan-2-yl)pyridin-3-yl)-2-methoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 HMz): δ=10.63 (1H, brs), 8.63 (1H, d), 8.19 (1H,d), 7.84 (1H, d), 7.76-7.65 (3H, m), 7.24 (1H, d), 7.07 (1H, d), 6.64(1H, dd), 3.85 (3H, s). MS: m/z 364.9 (M+H⁺).

Example III-70N-(5-(Furan-3-yl)pyridin-3-yl)-2,5-dimethoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.36 (1H, brs), 8.51 (1H, d), 8.22 (1H,s), 8.16 (1H, d), 7.80 (1H, s), 7.62 (1H, t), 7.31 (1H, d), 7.19-7.10(2H, m), 6.88 (1H, s), 3.80 (3H, s), 3.72 (3H, s). MS: m/z 361.0 (M+H⁺).

Example III-715-Bromo-N-(5-(furan-3-yl)pyridin-3-yl)-2-methoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.51 (1H, brs), 8.54 (1H, s), 8.24 (1H,s), 8.15 (1H, d), 7.87 (1H, d), 7.81-7.60 (2H, m), 7.61 (1H, s), 7.18(1H, d), 6.90 (1H, s), 3.85 (3H, s). MS: m/z 408.9 (M+H⁺).

Example III-725-Chloro-N-(5-(furan-3-yl)pyridin-3-yl)-2-methoxybenzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.51 (1H, brs), 8.54 (1H, d), 8.24 (1H,s), 8.16 (1H, d), 7.81-7.75 (2H, m), 7.68-7.61 (2H, m), 7.24 (1H, d),6.90 (1H, d), 3.86 (3H, s). MS: m/z 364.9 (M+H⁺).

Example III-732,5-Dimethoxy-N-(5-(thiophen-2-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.51 (1H, brs), 8.54 (1H, d), 8.24 (1H,s), 8.16 (1H, d), 7.81-7.75 (2H, m), 7.68-7.61 (2H, m), 7.24 (1H, d),6.90 (1H, d), 3.86 (3H, s). MS: m/z 364.9 (M+H⁺).

Example III-745-Bromo-2-methoxy-N-(5-(thiophen-2-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.64 (1H, brs), 8.60 (1H, d), 8.22 (1H,d), 7.89 (1H, d), 7.79 (1H, dd), 7.68-7.62 (2H, m), 7.56 (1H, dd),7.20-7.17 (2H, m), 3.86 (3H, s). MS: m/z 424.8 (M+H⁺).

Example III-755-Chloro-2-methoxy-N-(5-(thiophen-2-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.63 (1H, brs), 8.60 (1H, d), 8.22 (1H,d), 7.78 (1H, d), 7.69-7.64 (3H, m), 7.55 (1H, dd), 7.24 (1H, d), 7.18(1H, dd), 3.86 (3H, s). MS: m/z 380.9 (M+H⁺).

Example III-762,5-Dimethoxy-N-(5-(thiophen-3-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.39 (1H, brs), 8.60 (1H, s), 8.21 (1H,s), 7.91 (1H, s), 7.73-7.68 (2H, m), 7.45 (1H, d), 7.31 (1H, d),7.16-7.13 (2H, m), 3.80 (3H, s), 3.71 (3H, s). MS: m/z 377.0 (M+H⁺).

Example III-775-Bromo-2-methoxy-N-(5-(thiophen-3-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.54 (1H, brs), 8.63 (1H, s), 8.21 (1H,s), 7.93 (1H, s), 7.87 (1H, d), 7.80-7.72 (3H, m), 7.47 (1H, d), 7.18(1H, d), 3.86 (3H, s). MS: m/z 424.9 (M+H⁺).

Example III-785-Chloro-2-methoxy-N-(5-(thiophen-3-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.55 (1H, brs), 8.63 (1H, s), 8.21 (1H,s), 7.93 (1H, d), 7.78-7.63 (4H, m), 7.47 (1H, d), 7.23 (1H, d), 3.87(3H, s). MS: m/z 380.9 (M+H⁺).

Example III-792,5-Dimethoxy-N-(5-(thiazol-2-yl)pyridin-3-yl)benzenesulfonamide

Step 1:5-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridin-3-ylamine (1.25g, 5.7 mmol), 2-bromo-thiazole (923 mg, 5.7 mmol), Cs₂CO₃ (7.4 g, 22.8mmol), Pd(PPh₃)₄ (200 mg) were stirred in 1,4-dioxane (15 mL) and water(3 mL) at 100° C. under N₂ for 4 h. Then the solvent was concentratedunder reduced-pressure. The residue was dissolved in water and theaqueous phase was extracted with EtOAc (30 mL×3). The organic layer waswashed with brine, dried over anhydrous Na₂SO₄ and concentrated todryness under reduced pressure. The crude was purified via silica gelcolumn (DCM/MeOH, 20/1) to afford 350 mg (yield: 35%) of5-thiazol-2-yl-pyridin-3-ylamine. MS: m/z 177.9 (M+H⁺).

Step 2: This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d₆, 400 HMz): δ=10.65 (1H, brs), 8.87 (1H, d), 8.39 (1H,d), 8.06 (1H, t), 7.99 (1H, d), 7.89 (1H, d), 7.34 (1H, d), 7.17-7.11(2H, m), 3.79 (3H, s), 3.78 (3H, s). MS: m/z 378.0 (M+H⁺).

Example III-805-Bromo-2-methoxy-N-(5-(thiazol-2-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d₆, 400 HMz): δ=10.76 (1H, brs), 8.78 (1H, s), 8.40 (1H,s), 8.05-8.00 (2H, m), 7.91-7.87 (2H, m), 7.79-7.77 (1H, m), 7.18 (1H,d), 3.84 (3H, s). MS: m/z 425.9 (M+H⁺).

Example III-815-Chloro-2-methoxy-N-(5-(thiazol-2-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO-d₆, 400 HMz): δ=10.78 (1H, brs), 8.79 (1H, s), 8.41 (1H,s), 8.07-8.01 (2H, m), 7.92 (1H, s), 7.80 (1H, s), 7.69-7.67 (1H, m),7.25 (1H, d), 3.86 (3H, s). MS: m/z 382.0 (M+H⁺).

Example III-822,5-Dimethoxy-N-(5-(thiazol-5-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example 111-79.

¹H NMR (DMSO-d₆, 400 HMz): δ=10.45 (1H, brs), 9.24 (1H, d), 8.81 (1H,d), 8.29-8.27 (2H, m), 8.11 (1H, t), 7.31 (1H, d), 7.17-7.12 (2H, m),3.80 (3H, s), 3.72 (3H, s). MS: m/z 378.0 (M+H⁺).

Example III-835-Bromo-2-methoxy-N-(5-(thiazol-5-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-79.

¹H NMR (DMSO-d₆, 400 HMz): δ=10.58 (1H, brs), 9.25 (1H, d), 8.85 (1H,s), 8.31 (1H, s), 8.27 (1H, d), 8.10 (1H, s), 7.85 (1H, d), 7.78-7.75(1H, m), 7.17 (1H, d), 3.85 (3H, s). MS: m/z 425.9 (M+H⁺).

Example III-845-Chloro-2-methoxy-N-(5-(thiazol-5-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-79.

¹H NMR (DMSO-d₆, 400 HMz): δ=10.59 (1H, brs), 9.25 (1H, d), 8.85 (1H,d), 8.31 (1H, d), 8.27 (1H, d), 8.11 (1H, t), 7.75 (1H, d), 7.65 (1H,dd), 7.23 (1H, d), 3.86 (3H, s). MS: m/z 382.0 (M+H⁺).

Example III-852,5-Dimethoxy-N-(5-(thiazol-4-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-79.

¹H NMR (DMSO-d₆, 400 HMz): δ=10.53 (1H, brs), 9.17 (1H, s), 8.60 (1H,s), 8.33 (1H, s), 8.27 (1H, d), 7.66 (1H, s), 7.33 (1H, d), 7.18-7.10(2H, m), 3.79 (3H, s), 3.73 (3H, s). MS: m/z 378.0 (M+H⁺).

Example III-865-Bromo-2-methoxy-N-(5-(thiazol-4-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-79.

¹H NMR (DMSO-d₆, 400 HMz): δ=10.67 (1H, brs), 9.19 (1H, s), 8.64 (1H,d), 8.35 (1H, s), 8.27 (1H, d), 7.89 (1H, d), 7.80 (1H, dd), 7.66 (1H,t), 7.18 (1H, d), 3.85 (3H, s). MS: m/z 425.9 (M+H⁺).

Example III-875-Chloro-2-methoxy-N-(5-(thiazol-4-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-79.

¹H NMR (DMSO-d₆, 400 HMz): δ=10.67 (1H, brs), 9.18 (1H, s), 8.63 (1H,s), 8.34 (1H, d), 8.27 (1H, d), 7.79 (1H, d), 7.69-7.65 (2H, m), 7.23(1H, d), 3.85 (3H, s). MS: m/z 382.0 (M+H⁺).

Example III-882,5-Dimethoxy-N-(5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO, 400 MHz): δ=10.30 (1H, brs), 8.45 (1H, d), 8.16 (1H, s),8.10 (1H, d), 7.80 (1H, s), 7.58 (1H, t), 7.30 (1H, d), 7.15-7.11 (2H,m), 3.86 (3H, s), 3.80 (3H, s), 3.71 (3H, s). MS: m/z 375.1 (M+H⁺).

Example III-895-Bromo-2-methoxy-N-(5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO, 400 MHz): δ=10.45 (1H, brs), 8.49 (1H, d), 8.17 (1H, s),8.09 (1H, d), 7.87-7.76 (3H, m), 7.57 (1H, t), 7.18 (1H, d), 3.87 (3H,s), 3.86 (3H, s). MS: m/z 423.0 (M+H⁺).

Example III-905-Chloro-2-methoxy-N-(5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-1.

¹H NMR (DMSO, 400 MHz): δ=10.45 (1H, brs), 8.49 (1H, d), 8.17 (1H, s),8.09 (1H, d), 7.82 (1H, s), 7.66 (1H, d), 7.65 (1H, dd), 7.57 (1H, t),7.23 (1H, d), 3.87 (3H, s). MS: m/z 379.0 (M+H⁺).

Example III-912,5-Dimethoxy-N-(5-(pyrimidin-2-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-79.

¹H NMR (DMSO-d6, 400 MHz): δ=10.60 (1H, brs), 9.14 (1H, s), 8.93 (2H,d), 8.50-8.43 (2H, m), 7.52 (1H, t), 7.33 (1H, d), 7.16-7.12 (2H, m),3.79 (3H, s), 3.73 (3H, s). MS: m/z 373.0 (M+H⁺).

Example III-925-Bromo-2-methoxy-N-(5-(pyrimidin-2-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-79.

¹H NMR (DMSO-d6, 400 MHz): δ=10.71 (1H, brs), 9.17 (1H, d), 8.95 (2H,d), 8.48 (1H, t), 8.44 (1H, d), 7.87 (1H, d), 7.78-7.75 (1H, m), 7.53(1H, t), 7.17 (1H, d), 3.84 (3H, s). MS: m/z 421.0 (M+H⁺).

Example III-935-Chloro-2-methoxy-N-(5-(pyrimidin-2-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-79.

¹H NMR (DMSO-d6, 400 MHz): δ=10.72 (1H, brs), 9.17 (1H, d), 8.95 (2H,d), 8.48 (1H, t), 8.45 (1H, d), 7.76 (1H, d), 7.65 (1H, dd), 7.53 (1H,t), 7.23 (1H, d), 3.85 (3H, s). MS: m/z 377.0 (M+H⁺).

Example III-942,5-Dimethoxy-N-(5-(pyrazin-2-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-79.

¹H NMR (DMSO-d6, 400 MHz): δ=10.59 (1H, brs), 9.26 (1H, s), 8.97 (1H,s), 8.77 (1H, s), 8.69 (1H, s), 8.43 (1H, s), 8.25 (1H, s), 7.34 (1H,s), 7.12-7.18 (2H, m), 3.80 (3H, s), 3.73 (3H, s). MS: m/z 373.0 (M+H⁺).

Example III-955-Bromo-2-methoxy-N-(5-(pyrazin-2-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-79.

¹H NMR (DMSO-d6, 400 MHz): δ=10.73 (1H, s), 9.28 (1H, s), 9.02 (1H, s),8.79-8.78 (1H, m), 8.70 (1H, s), 8.43 (1H, s), 8.24 (1H, s), 7.89 (1H,s), 7.77-7.80 (1H, m), 7.17 (1H, d), 3.85 (3H, s). MS: m/z 421.0 (M+H⁺).

Example III-965-Chloro-2-methoxy-N-(5-(pyrazin-2-yl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example III-79.

¹H NMR (DMSO-d6, 400 MHz): δ=10.74 (1H, brs), 9.29 (1H, s), 9.02 (1H,s), 8.79-8.70 (2H, m), 8.45 (1H, s), 8.26 (1H, s), 7.66-7.80 (2H, m),7.24-7.26 (1H, m), 3.87 (3H, s). MS: m/z 377.0 (M+H⁺).

Example IV Example IV-1 Methyl5-(5-bromo-2-methoxyphenylsulfonamido)nicotinate

To a solution of methyl 5-aminonicotinate (100 mg, 0.66 mmol) inpyridine (5 mL) was added 5-bromo-2-methoxybenzene-1-sulfonyl chloride(186 mg, 0.66 mmol) and DMAP (10 mg, 0.08 mmol), and the mixture wasthen heated at 60° C. overnight. LC-MS showed the reaction was complete.The resultant mixture was concentrated in vacuum and the residue wastriturated with methanol by ultrasonic-wave to give 87 mg (yield: 33%)of methyl 5-(5-bromo-2-methoxyphenylsulfonamido) nicotinate as a whitesolid.

¹H NMR (DMSO-d6): δ=10.82 (1H, brs), 8.74 (1H, d), 8.52 (1H, d), 8.00(1H, d), 7.87 (1H, d), 7.80 (1H, d), 7.18 (1H, d), 3.86 (3H, s), 3.80(3H, s). MS: m/z 401.0 (M+H⁺).

Example IV-2 Ethyl 5-(5-bromo-2-methoxyphenylsulfonamido)nicotinate

¹H NMR (DMSO-d6): δ=10.82 (1H, brs), 8.73 (1H, s), 8.53 (1H, s), 7.98(1H, s), 7.87 (1H, s), 7.80 (1H, d), 7.18 (1H, d), 4.32 (2H, q), 3.81(3H, s), 1.31 (3H, t). MS: m/z 414.9 (M+H⁺).

Example IV-3 Propyl 5-(5-bromo-2-methoxyphenylsulfonamido)nicotinate

¹H NMR (DMSO-d6): δ=10.83 (1H, brs), 8.73 (1H, d), 8.54 (1H, d), 7.99(1H, s), 7.87 (1H, d), 7.79 (1H, dd), 7.18 (1H, d), 4.25-4.21 (2H, m),3.82 (3H, s), 1.74-1.68 (2H, m), 0.96 (2H, t). MS: m/z 429.0 (M+H⁺).

Example IV-4 Cyclohexyl 5-(5-bromo-2-methoxyphenylsulfonamido)nicotinate

Step 1: To a solution of methyl5-(5-bromo-2-methoxyphenylsulfonamido)nicotinate (1 g, 2.5 mmol) in THF(20 mL) was added aq. NaOH (2M, 10 mL), it was then heated at 60° C. for3 h. TLC showed that the reaction was complete. The solution wasconcentrated in vacuum to remove THF. The remaining aqueous phase wasadjusted pidified to pH=2 with 2N HCl. The resulting solid was filteredto give 0.95 g (yield: 98%) of5-(5-bromo-2-methoxyphenylsulfonamido)nicotinic acid as a white solid.

Step 2: To a solution of 5-(5-bromo-2-methoxyphenylsulfonamido)nicotinicacid (80 mg, 0.21 mmol) in cyclohexanol (5 mL) was added SOCl₂ (0.2 mL),it was then refluxed overnight. LC-MS showed the reaction was complete.The resultant was concentrated in vacuum to remove cyclohexanol. Theresidue was re-crystallized from methanol to give 54 mg (yield: 55%) ofcyclohexyl 5-(5-bromo-2-methoxyphenylsulfonamido) nicotinate as a whitesolid.

¹H NMR (DMSO-d6): δ=10.84 (1H, brs), 8.73 (1H, d), 8.54 (1H, d), 7.98(1H, t), 7.87 (1H, d), 7.81 (1H, dd), 7.18 (1H, d), 4.96-4.92 (1H, m),3.83 (3H, s), 1.84-1.62 (4H, m), 1.58-1.34 (6H, m). MS: m/z 469.0 (M+H⁺)

Example IV-5 Phenyl 5-(5-bromo-2-methoxyphenylsulfonamido)nicotinate

¹H NMR (DMSO-d6): δ=10.90 (1H, brs), 8.93 (1H, s), 8.61 (1H, d), 8.13(1H, s), 7.89 (1H, d), 7.82 (1H, d), 7.48 (2H, t), 7.36-7.30 (3H, m),7.20 (1H, d), 3.84 (3H, s). MS: m/z 463.0 (M+H⁺)

Example IV-6 Methyl 5-(5-chloro-2-methoxyphenylsulfonamido)nicotinate

1H NMR (DMSO-d6): δ=10.82 (1H, brs), 8.73 (1H, s), 8.54 (1H, d), 7.99(1H, s), 7.77 (1H, d), 7.68 (1H, dd), 7.23 (1H, d), 3.86 (3H, s), 3.82(3H, s). MS: m/z 356.1 (M+H⁺).

Example IV-7 Ethyl 5-(5-chloro-2-methoxyphenylsulfonamido)nicotinate

¹H NMR (DMSO-d6): δ=10.83 (1H, brs), 8.74 (1H, d), 8.52 (1H, d), 7.98(1H, s), 7.77 (1H, d), 7.68 (1H, dd), 7.23 (1H, d), 4.33-4.29 (2H, m),3.82 (3H, s), 1.31 (2H, t). MS: m/z 371.0 (M+H⁺).

Example IV-8 Propyl 5-(5-chloro-2-methoxyphenylsulfonamido)nicotinate

¹H NMR (DMSO-d6): δ=10.84 (1H, brs), 8.74 (1H, d), 8.54 (1H, d), 7.99(1H, t), 7.77 (1H, d), 7.69 (1H, dd), 7.23 (1H, d), 4.23 (2H, t), 3.82(3H, s), 1.75-1.67 (2H, m), 0.96 (2H, t). MS: m/z 385.0 (M+H⁺).

Example IV-9 Cyclohexyl5-(5-chloro-2-methoxyphenylsulfonamido)nicotinate

¹H NMR (DMSO-d6): δ=10.84 (1H, brs), 8.74-8.73 (1H, d), 8.55-8.54 (1H,d), 7.99 (1H, s), 7.78-7.77 (1H, d), 7.71 (1H, dd), 7.26-7.24 (1H, d),4.97-4.93 (1H, m), 3.84 (3H, s), 1.86-1.83 (2H, m), 1.73-1.67 (2H, m),1.58-1.35 (6H, m). MS: m/z 425.1 (M+H⁺).

Example IV-10 Phenyl 5-(5-chloro-2-methoxyphenylsulfonamido)nicotinate

¹H NMR (DMSO-d6): δ=10.90 (1H, brs), 8.92 (1H, s), 8.62 (1H, d), 8.13(1H, s), 7.80 (1H, d), 7.71 (1H, dd), 7.52-7.45 (2H, m), 7.35-7.25 (4H,m), 3.85 (3H, s). MS: m/z 419.0 (M+H⁺).

Example IV-11 Methyl 5-(2,5-dimethoxyphenylsulfonamido)nicotinate

¹H NMR (DMSO-d6): δ=10.70 (1H, brs), 8.71 (1H, d), 8.53 (1H, d), 8.01(1H, t), 7.32 (1H, d), 7.22-7.12 (2H, m), 3.86 (3H, s), 3.76 (3H, s),3.74 (3H, s). MS: m/z 353.1 (M+H⁺).

Example IV-12 Ethyl 5-(2,5-dimethoxyphenylsulfonamido)nicotinate

¹H NMR (DMSO-d6): δ=10.70 (1H, brs), 8.71 (1H, d), 8.54 (1H, d), 7.99(1H, t), 7.32 (1H, d), 7.19-7.12 (2H, m), 4.32 (2H, q), 3.77 (3H, s),3.74 (3H, s), 1.31 (3H, t). MS: m/z 367.1 (M+H⁺).

Example IV-13 Propyl 5-(2,5-dimethoxyphenylsulfonamido)nicotinate

¹H NMR (DMSO-d6): δ=10.69 (1H, s), 8.70 (1H, s), 8.53-8.52 (1H, d), 7.99(1H, s), 7.31-7.30 (1H, d), 7.20-7.12 (2H, m), 4.24-4.21 (2H, t), 3.76(3H, s), 3.73 (3H, s), 1.72-1.67 (2H, m), 0.96-0.92 (3H, t) ppm MS: m/z381 (M+H+)

Example IV-14 Cyclohexyl 5-(2,5-dimethoxyphenylsulfonamido)nicotinate

¹H NMR (DMSO-d6): δ=10.69 (1H, brs), 8.70 (1H, s), 8.54 (1H, d), 7.99(1H, s), 7.33 (1H, d), 7.19-7.15 (2H, m), 4.97-4.92 (1H, m), 3.78 (3H,s), 3.74 (3H, s), 1.88-1.82 (2H, m), 1.72-1.64 (2H, m), 1.55-1.36 (6H,m). MS: m/z 421.2 (M+H⁺).

Example IV-15 Phenyl 5-(2,5-dimethoxyphenylsulfonamido)nicotinate

¹H NMR (DMSO-d6): δ=10.78 (1H, brs), 8.89 (1H, d), 8.61 (1H, d), 8.13(1H, d), 7.48 (2H, t), 7.35-7.27 (4H, m), 7.22-7.15 (2H, m), 3.79 (3H,s), 3.73 (3H, s). MS: m/z 415.1 (M+H⁺).

Example IV-165-(5-Bromo-2-methoxyphenylsulfonamido)-N-methylnicotinamide

Step 1: To a stirred solution of 5-amino-nicotinic acid (10.0 g, 72.5mmol) in methanol (100 mL) was added SOCl₂ (10.4 g, 86.9 mmol) dropwiseat 0° C. The mixture was allowed warm to room temperature and thenrefluxed for 16 hours. The mixture was cooled, concentrated in vacuumand the residue was diluted with water (200 mL). The mixture wasneutralized with aqueous NaHCO₃ solution to pH=7. The aqueous mixturewas extracted with DCM (100 mL×2). The combined organic layers werewashed with brine (100 mL×2), dried over anhydrous Na₂SO₄, filtered andthe filtrate was concentrated in vacuum to dryness to give 9.5 g (yield:86%) of 5-amino-nicotinic acid methyl ester as white solid.

¹H NMR (DMSO-d₆, 400 MHz): δ=8.24 (1H, d), 8.12 (1H, d), 7.42 (1H, dd),5.65 (2H, brs), 3.84 (3H, s).

Step 2: To a stirred HSO₃Cl (100 g) was added 1-bromo-4-methoxy-benzene(15.0 g, 80.6 mmol) dropwise at 25° C. The mixture was stirred at thistemperature for 16 hours. The mixture was poured into ice water (1 L)dropwise and the resulting solid was filtered. The solid was evaporatedin vacuum to dryness to give 17.3 g (yield: 75%) of5-bromo-2-methoxy-benzenesulfonyl chloride as white solid.

¹H NMR (DMSO-d₆, 400 MHz): δ=7.77 (1H, d), 7.47 (1H, dd), 6.96 (1H, d),3.76 (3H, s).

Step 3: To a stirred mixture of 5-amino-nicotinic acid methyl ester (6.5g, 42.8 mmol) and 5-bromo-2-methoxy-benzenesulfonyl chloride (15.8 g,55.6 mmol) in pyridine (60 mL) was added DMAP (260 mg, 2.14 mmol). Themixture was stirred at 80° C. for 17 hours. The mixture was cooled,concentrated in vacuum to dryness. The residue was diluted with MeOH(100 mL) and stirred for 30 minutes. The suspended solid was filteredand washed with methanol (50 mL), evaporated in vacuum to dryness togive 12.1 g (yield: 70%) of5-(5-bromo-2-methoxy-benzenesulfonylamino)-nicotinic acid methyl esteras white solid.

¹H NMR (DMSO-d₆, 400 MHz): δ=10.86 (1H, brs), 8.80 (1H, s), 8.59 (1H,d), 8.06 (1H, s), 7.93 (1H, d), 7.85 (1H, dd), 7.24 (1H, d), 3.92 (3H,s), 3.86 (3H, s).

Step 4: To a solution of methylamine ethanol solution (30-33%, 30 mL)was added 5-(5-bromo-2-methoxy-benzenesulfonylamino)-nicotinic acidmethyl ester (2.0 g, 5 mmol). The mixture was stirred at 80° C. for 17hours, cooled, and concentrated in vacuum to dryness. The residue waspurified by silica gel chromatography (from DCM to DCM/MeOH=20/1) togive white solid. The solid was washed with methanol (10 mL) andevaporated in vacuum to dryness to give 1.2 g (yield: 60%) of5-(5-bromo-2-methoxy-benzenesulfonylamino)-N-methyl-nicotinamide aswhite solid.

¹H NMR (DMSO-d₆, 400 MHz): δ=10.62 (1H, brs), 8.61-8.65 (2H, m), 8.40(1H, d), 7.89 (1H, dd), 7.83 (1H, d), 7.77 (1H, dd), 7.17 (1H, d), 3.82(3H, s), 2.77 (3H, d). MS: m/z 400.0 (M+H⁺).

Example IV-17 5-(5-Bromo-2-methoxy-benzenesulfonylamino)-nicotinamide

To a solution of ammonium hydroxide (28-29%, 60 mL) was added5-(5-bromo-2-methoxy-benzenesulfonylamino)-nicotinic acid methyl ester(10.0 g, 25 mmol). The mixture was stirred at 80° C. for 16 hours,cooled, and concentrated to give white solid. The solid was washed withmethanol (20 mL×2) and evaporated in vacuum to dryness to give 8.2 g(yield: 85%) of 5-(5-bromo-2-methoxy-benzenesulfonylamino)-nicotinamideas white solid.

¹H NMR (DMSO-d₆, 400 MHz): δ=10.62 (1H, brs), 8.70 (1H, d), 8.41 (1H,d), 8.13 (1H, brs), 7.91 (1H, s), 7.83 (1H, d), 7.77 (1H, dd), 7.61 (1H,brs), 7.17 (1H, d), 3.82 (3H, s). MS: m/z 385.9 (M+H⁺).

Example IV-18 5-(5-Bromo-2-methoxyphenylsulfonamido)-N-ethylnicotinamide

¹H NMR (DMSO-d6): δ=10.65 (1H, brs), 8.68-8.64 (2H, m), 8.40 (1H, d),7.88 (1H, s), 7.84 (1H, s), 7.78 (1H, d), 7.18 (1H, d), 3.82 (3H, s),3.27-3.24 (2H, m), 1.12-1.08 (3H, t). MS: m/z 414.1 (M+H⁺).

Example IV-195-(5-Bromo-2-methoxyphenylsulfonamido)-N-propylnicotinamide

¹H NMR (DMSO-d6): δ=10.64 (1H, brs), 8.66-8.63 (2H, m), 8.47 (1H, s),7.87-7.83 (3H, m), 7.18 (1H, s), 3.81 (3H, s), 3.21-3.17 (2H, m),1.53-1.49 (2H, m), 0.89-0.82 (3H, m). MS: m/z 427.1 (M+H⁺)

Example IV-205-(5-Bromo-2-methoxyphenylsulfonamido)-N-cyclohexylnicotinamide

To a solution of 5-(5-bromo-2-methoxyphenylsulfonamido)nicotinic acid(80 mg, 0.21 mmol) in DMF (5 mL) was added DIEA (21 mg, 0.21 mmol),cyclohexylamine (20 mg, 0.21 mmol) and HATU (80 mg, 0.21 mmol), it wasthen stirred at room temperature for 4 h. LC-MS showed the reaction wascomplete. The resultant was concentrated in vacuum to remove most of DMFand the residue was re-crystallized from methanol to give 50 mg (yield:51%) of 5-(5-bromo-2-methoxyphenylsulfonamido)-N-cyclohexylnicotinamideas a white solid.

¹H NMR (DMSO-d6): δ=10.62 (1H, brs), 8.66 (1H, s), 8.44-8.38 (2H, m),7.86 (1H, s), 7.83 (1H, d), 7.78 (1H, dd), 7.18 (1H, d), 3.82 (3H, s),3.70-3.81 (1H, m) 1.80-1.57 (5H, m), 1.32-1.10 (5H, m). MS: m/z 468.1(M+H⁺)

Example IV-215-(5-Bromo-2-methoxyphenylsulfonamido)-N-(2-methoxyethyl)nicotinamide

¹H NMR (DMSO-d6): δ=10.65 (1H, brs), 8.75 (1H, s), 8.67 (1H, s), 8.40(1H, d), 7.89 (1H, s), 7.84 (1H, d), 7.79 (1H, dd), 7.17 (1H, d), 3.81(3H, s), 3.39-3.44 (4H, m), 3.22 (3H, s). MS: m/z 444.0 (M+H⁺)

Example IV-225-(5-Bromo-2-methoxyphenylsulfonamido)-N-(2-(dimethylamino)ethyl)nicotinamide

¹H NMR (DMSO-d6): δ=10.72 (1H, brs), 9.42 (1H, brs, TFA salt), 8.91 (1H,t), 8.72 (1H, s), 8.43 (1H, d), 7.94 (1H, s), 7.84 (1H, d), 7.79 (1H,dd), 7.19 (1H, d), 3.84 (3H, s), 3.61-3.58 (2H, m), 3.27-3.23 (2H, m),2.84 (3H, s), 2.83 (3H, s). MS: m/z 457.1 (M+H⁺).

Example IV-235-(5-Bromo-2-methoxyphenylsulfonamido)-N-phenylnicotinamide

¹H NMR (DMSO-d6): δ=10.73 (1H, brs), 10.45 (1H, brs), 8.81 (1H, s), 8.48(1H, d), 7.95 (1H, s), 7.86 (1H, d), 7.79 (1H, dd), 7.72 (2H, d), 7.36(2H, t), 7.16 (1H, d), 7.13 (1H, t), 3.84 (3H, s). MS: m/z 462.0 (M+H⁺).

Example IV-245-Bromo-2-methoxy-N-(5-(morpholine-4-carbonyl)pyridin-3-yl)benzenesulfonamide

¹H NMR (DMSO-d6): δ=10.70 (1H, brs), 8.36 (1H, d), 8.28 (1H, s), 7.86(1H, d), 7.79 (1H, dd), 7.48 (1H, s), 7.17 (1H, d), 3.84 (3H, s),3.70-3.50 (6H, m), 3.17-3.12 (2H, m). MS: m/z 456.0 (M+H⁺)

Example IV-255-Bromo-2-methoxy-N-(5-(4-methylpiperazine-1-carbonyl)pyridin-3-yl)benzenesulfonamide

¹H NMR (DMSO-d6): δ=10.71 (1H, brs), 8.35 (1H, s), 8.24 (1H, s), 7.84(1H, d), 7.79 (1H, dd), 7.44 (1H, s), 7.17 (1H, d), 3.84 (3H, s),3.60-3.56 (2H, m), 3.16-3.10 (2H, m), 2.37-2.34 (2H, m), 2.33-2.18 (5H,s). MS: m/z 469.0 (M+H⁺)

Example IV-26 5-(5-Chloro-2-methoxyphenylsulfonamido)nicotinamide

¹H NMR (DMSO-d6): δ=10.65 (1H, brs), 8.70 (1H, d), 8.42 (1H, d), 8.15(1H, s), 7.91 (1H, t), 7.74 (1H, d), 7.68-7.63 (2H, m), 7.23 (1H, d),3.83 (3H, s). MS: m/z 342.0 (M+H⁺).

Example IV-275-(5-Chloro-2-methoxyphenylsulfonamido)-N-methylnicotinamide

¹H NMR (DMSO-d6): δ=10.65 (1H, brs), 8.67-8.62 (2H, m), 8.41 (1H, s),7.89 (1H, s), 7.73 (1H, s), 7.74-7.66 (1H, m), 7.24-7.22 (1H, m), 3.83(3H, s), 2.77 (3H, s). MS: m/z 356.0 (M+H⁺)

Example IV-285-(5-Chloro-2-methoxyphenylsulfonamido)-N-ethylnicotinamide

¹H NMR (DMSO-d6): δ=10.64 (1H, brs), 8.67-8.65 (2H, m), 8.41 (1H, d),7.88 (1H, s), 7.74 (1H, d), 7.69 (1H, dd), 7.23 (1H, d), 3.83 (3H, s),3.28-3.25 (2H, m), 1.10 (3H, t). MS: m/z 370.1 (M+H⁺)

Example IV-295-(5-Chloro-2-methoxyphenylsulfonamido)-N-propylnicotinamide

¹H NMR (DMSO-d6): δ=10.66 (1H, brs), 8.66-8.63 (2H, m), 8.39 (1H, d),7.87 (1H, t), 7.73 (1H, d), 7.66 (1H, d), 7.23 (1H, d), 3.83 (3H, s),3.20 (2H, q), 1.52-1.50 (2H, m), 0.875 (3H, t). MS: m/z 384.1 (M+H⁺).

Example IV-305-(5-Chloro-2-methoxyphenylsulfonamido)-N-cyclohexylnicotinamide

¹H NMR (DMSO-d6): δ=10.63 (1H, brs), 8.65 (1H, s), 8.42-8.38 (2H, m),7.85 (1H, d), 7.72 (1H, d), 7.68 (1H, dd), 7.23 (1H, d), 3.83 (3H, s),3.74-3.71 (1H, m) 1.80-1.71 (5H, m), 1.30-1.10 (5H, m). MS: m/z 424.1(M+H⁺)

Example IV-315-(5-Chloro-2-methoxyphenylsulfonamido)-N-phenylnicotinamide

¹H NMR (DMSO-d6): δ=10.73 (1H, brs), 10.45 (1H, s), 8.81 (1H, s), 8.49(1H, s), 7.96 (1H, s), 7.77-7.68 (4H, m), 7.39-7.35 (2H, t), 7.27-7.24(1H, d), 7.15-7.11 (1H, m), 3.86 (3H, s). MS: m/z 418.1 (M+H⁺)

Example IV-325-(5-Chloro-2-methyoxyphenylsulfonamido)-N-(2-methoxyethyl)nicotinamide

¹H NMR (DMSO-d6): δ=10.66 (1H, brs), 8.75-8.73 (1H, m), 8.67 (1H, d),8.41 (1H, d), 7.90 (1H, t), 7.74 (1H, d), 7.68 (1H, dd), 7.23 (1H, d),3.83 (3H, s), 3.44-3.40 (4H, m), 3.26 (3H, s). MS: m/z 400.1 (M+H⁺).

Example IV-335-(5-Chloro-2-methoxyphenylsulfonamido)-N-(2-(dimethylamino)ethyl)nicotinamide

¹H NMR (DMSO-d6): δ=10.73 (1H, brs), 9.56 (1H, brs, TFA salt), 8.93-8.91(1H, m), 8.72 (1H, s), 8.45 (1H, d), 7.95 (1H, s), 7.75 (1H, d), 7.69(1H, dd), 7.25 (1H, d), 3.85 (3H, s), 3.62-3.58 (2H, m), 3.27-3.25 (2H,m), 2.85 (3H, s), 2.84 (3H, s). MS: m/z 413.1 (M+H⁺)

Example IV-345-Chloro-2-methoxy-N-(5-(morpholine-4-carbonyl)pyridin-3-yl)benzenesulfonamide

¹H NMR (DMSO-d6): δ=10.70 (1H, brs), 8.37 (1H, d), 8.28 (1H, d), 7.76(1H, d), 7.69 (1H, dd), 7.48 (1H, t), 7.23 (1H, d), 3.84 (3H, s),3.75-3.47 (6H, m), 3.18-3.12 (2H, m). MS: m/z 412.1 (M+H⁺).

Example IV-355-Chloro-2-methoxy-N-(5-(4-methylpiperazine-1-carbonyl)pyridin-3-yl)benzenesulfonamide

¹H NMR (DMSO-d6): δ=10.77 (1H, brs), 10.18 (1H, brs, TFA salt), 8.38(1H, s), 8.25 (1H, s), 7.76 (1H, d), 7.69 (1H, dd), 7.61 (1H, d), 7.24(1H, d), 3.83 (3H, s), 3.81-3.00 (8H, m), 2.82 (3H, s). MS: m/z 425.1(M+H⁺).

Example IV-36 5-(2,5-Dimethoxyphenylsulfonamido)nicotinamide

¹H NMR (DMSO-d6): δ=10.51 (1H, brs), 8.66 (1H, s), 8.40 (1H, d), 8.13(1H, s), 7.91 (1H, s), 7.61 (1H, s), 7.28 (1H, d), 7.16-7.11 (2H, m),3.77 (3H, s), 3.73 (3H, s). MS: m/z 338.1 (M+H⁺).

Example IV-37 5-(2,5-Dimethoxyphenylsulfonamido)-N-methylnicotinamide

¹H NMR (DMSO-d6): δ=10.51 (1H, brs), 8.63-8.59 (2H, m), 8.40 (1H, d),7.89 (1H, s), 7.28 (1H, d), 7.17-7.12 (2H, m), 3.77 (3H, s), 3.72 (3H,s), 2.75 (3H, d). MS: m/z 352.1 (M+H⁺)

Example IV-38 5-(2,5-Dimethoxyphenylsulfonamido)-N-propylnicotinamide

¹H NMR (DMSO-d6): δ=10.53 (1H, brs), 8.64-8.61 (2H, m), 8.40 (1H, d),7.88 (1H, s), 7.28 (1H, d), 7.19-7.10 (2H, m), 3.77 (3H, s), 3.73 (3H,s), 3.18 (2H, q), 1.55-1.46 (2H, m), 0.87 (3H, t). MS: m/z 380.1 (M+H⁺)

Example IV-395-(2,5-Dimethoxyphenylsulfonamido)-N-cyclohexylnicotinamide

¹H NMR (DMSO-d6): δ=10.49 (1H, brs), 8.62 (1H, s), 8.41-8.36 (2H, m),7.86 (1H, d), 7.28 (1H, d), 7.17-7.12 (2H, m), 3.77 (3H, s), 3.76-3.72(4H, m), 1.75-1.52 (5H, m), 1.29-1.00 (5H, m). MS: m/z 420.2 (M+H⁺).

Example IV-40 5-(2,5-Dimethoxyphenylsulfonamido)-N-phenylnicotinamide

¹H NMR (DMSO-d6): δ=10.60 (1H, brs), 10.43 (1H, brs), 8.77 (1H, s), 8.48(1H, d), 7.96 (1H, d), 7.72 (2H, m), 7.38-7.30 (3H, m), 7.19-7.11 (3H,m), 3.79 (3H, s), 3.73 (3H, s). MS: m/z 414.1 (M+H⁺)

Example IV-415-(2,5-Dimethoxyphenylsulfonamido)-N-(2-methoxyethyl)nicotinamide

¹H NMR (DMSO-d6): δ=10.51 (1H, brs), 8.72 (1H, s), 8.63 (1H, s), 8.40(1H, s), 7.89 (1H, s), 7.29 (1H, s), 7.13 (2H, m), 3.77 (3H, s), 3.72(3H, s), 3.43-3.39 (4H, m), 3.24 (3H, s). MS: m/z 396.2 (M+H⁺).

Example IV-425-(2,5-Dimethoxyphenylsulfonamido)-N-(2-(dimethylamino)ethyl)nicotinamide

¹H NMR (DMSO-d6): δ=10.59 (1H, brs), 9.35 (1H, brs, TFA salt), 8.86 (1H,s), 8.67 (1H, s), 8.43 (1H, d), 7.94 (1H, s), 7.29 (1H, d), 7.19-7.13(2H, m), 3.78 (3H, s), 3.73 (3H, s), 3.59-3.56 (2H, m), 3.26-3.22 (2H,m), 2.84 (3H, s), 2.83 (3H, s). MS: m/z 409.2 (M+H⁺)

Example IV-432,5-Dimethoxy-N-(5-(morpholine-4-carbonyl)pyridin-3-yl)benzenesulfonamide

¹H NMR (DMSO-d6): δ=10.56 (1H, brs), 8.37 (1H, d), 8.24 (1H, d), 7.47(1H, t), 7.29 (1H, d), 7.18-7.10 (2H, m), 3.78 (3H, s), 3.73 (3H, s),3.72-3.48 (6H, m), 3.45-3.10 (2H, m). MS: m/z 408.1 (M+H⁺).

Example IV-442,5-Dimethoxy-N-(5-(4-methylpiperazine-1-carbonyl)pyridin-3-yl)benzenesulfonamide

¹H NMR (DMSO-d6): δ=10.63 (1H, brs), 10.17 (1H, brs, TFA salt), 8.37(1H, d), 8.29 (1H, d), 7.60 (1H, t), 7.30 (1H, d), 7.20-7.13 (2H, m),3.77 (3H, s), 3.74 (3H, s), 3.61-3.00 (8H, m), 2.81 (3H, s). MS: m/z421.2 (M+H⁺).

Example V Example V-15-Bromo-2-methoxy-N-(thiophen-3-yl)benzenesulfonamide

The mixture of 5-bromo-2-methoxy-benzenesulfonyl chloride (145 mg, 0.51mmol), Thiophen-3-ylamine (50 mg, 0.51 mmol), DMAP (75 mg, 0.61 mmol) inpyridine (2 mL) was stirred at 60° C. for 4 h. And then, water (10 mL)was added and the reaction mixture was extracted with DCM (10 mL×3). Theextracts were dried over Na₂SO₄ and concentrated to dryness. The residuewas purified by prep-HPLC to afford 60 mg (yield: 36%) of5-bromo-2-methoxy-N-(thiophen-3-yl)benzenesulfonamide.

¹H NMR (DMSO-d6): δ=10.28 (1H, brs), 7.78-7.75 (2H, m), 7.38 (1H, dd),7.18 (1H, d), 6.86-6.83 (2H, m), 3.90 (3H, s). MS: m/z 348.0 (M+H⁺).

Example V-2 2,5-Dimethoxy-N-(thiophen-3-yl)benzenesulfonamide

¹H NMR (DMSO-d6): δ=10.15 (1H, s), 7.37-7.36 (1H, d), 7.25-7.24 (1H, d),7.15-7.13 (2H, m), 6.84-6.81 (2H, m), 3.84 (3H, s), 3.73 (3H, s). MS:m/z 300 (M+H⁺).

Example V-3 5-Bromo-2-methoxy-N-oxazol-2-yl-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.90 (1H, s), 7.89 (1H, d), 7.74 (1H, dd),7.66 (1H, d), 7.29 (1H, d), 7.16 (1H, d), 3.73 (3H, s). MS: m/z 334.9(M+H⁺).

Example V-4 2,5-Dimethoxy-N-oxazol-2-yl-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.76 (1H, s), 7.64 (1H, s), 7.35 (1H, d),7.27 (1H, s), 7.14-7.10 (2H, m), 3.77 (3H, s), 3.67 (3H, s). MS: m/z285.1 (M+H⁺).

Example V-55-Bromo-2-methoxy-N-(3-methyl-isoxazol-5-yl)-benzenesulfonamide

The mixture of 5-bromo-2-methoxy-benzenesulfonyl chloride (145 mg, 0.51mmol), 3-methyl-isoxazol-5-ylamine (50 mg, 0.51 mmol), DMAP (75 mg, 0.61mmol) in pyridine (2 mL) was stirred at 40° C. for 4 h. And then, water(10 mL) was added and the reaction mixture was extracted with DCM (10mL×3). The extracts were dried over Na₂SO₄ and concentrated to dryness.The residue was purified by prep-HPLC to afford 10 mg (yield: 6%) of5-bromo-2-methoxy-N-(3-methyl-isoxazol-5-yl)-benzenesulfonamide asyellow solid.

¹H NMR (DMSO-d6, 400 MHz): δ=12.13 (1H, s), 7.88 (1H, s), 7.85 (1H, d),7.26 (1H, d), 5.65 (1H, s), 3.87 (3H, s), 2.11 (3H, s). MS: m/z 348.9(M+H⁺).

Example V-6 2,5-Dimethoxy-N-(3-methyl-isoxazol-5-yl)-benzenesulfonamide

¹H NMR (DMSO-d6, 400 MHz): δ=11.99 (1H, brs), 7.32-7.17 (3H, m), 5.59(1H, s), 3.80 (3H, s), 3.77 (3H, s), 2.08 (3H, s). MS: m/z 298.9 (M+H⁺).

Example V-7 5-Bromo-2-methoxy-N-(1H-pyrazol-3-yl)-benzenesulfonamide

Step 1: To the mixture of 1H-pyrazol-3-ylamine (500 mg, 6.0 mmol), TEA(1.21 g, 12.0 mmol), DMAP (50 mg, 0.4 mmol) in dioxane (20 mL), wasadded (Boc)₂O (1.5 g, 6.9 mmol) dropwise at r.t. The mixture was stirredat r.t. for 4 h. The solution was concentrated in vacuum. The residuewas diluted with EtOAc (20 mL), washed with water (20 mL×2), brine (20mL) and dried over magnesium sulfate. The solution was filtered andconcentrated in vacuum to afford 700 mg (yield: 64%) of crude3-amino-pyrazole-1-carboxylic acid tert-butyl ester. MS: m/z 184.0(M+H⁺).

Step 2: This step is similar to Example V-1. MS: m/z 430.0 (M−H⁺).

Step 3: The mixture of3-(5-bromo-2-methoxy-benzenesulfonylamino)-pyrazole-1-carboxylic acidtert-butyl ester (200 mg, 0.46 mmol) in HCl (4.0 M in MeOH, 10 mL) wasstirred at r.t. for 60 minutes. The solvent was evaporated and theresidue was purified by prep-TLC (PE/EtOAc, 2/1) to afford 11 mg (yield:7%) of 5-bromo-2-methoxy-N-(1H-pyrazol-3-yl)-benzenesulfonamide as whitesolid.

¹H NMR (DMSO-d6, 400 MHz): δ=12.34 (1H, s), 7.78-7.73 (2H, m), 7.49 (1H,t), 7.17 (1H, d), 5.85 (1H, s), 3.85 (3H, s). MS: m/z 331.8 (M+H⁺).

Example V-8 2,5-Dimethoxy-N-(1H-pyrazol-3-yl)-benzenesulfonamide

This compound was prepared as described in Example V-5.

¹H NMR (DMSO-d6, 400 MHz): δ=7.96 (1H, d), 7.29-7.25 (2H, m), 7.16 (1H,d), 5.81 (1H, d), 5.50 (2H, s), 3.77 (3H, s), 3.70 (3H, s). MS: m/z284.0 (M+H⁺).

Example V-9 5-Bromo-2-methoxy-N-(1-methyl-1H-pyrazol-3-yl)-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.27 (1H, s), 7.76-7.73 (2H, m), 7.46 (1H,d), 7.18 (1H, d), 5.81 (1H, d), 3.87 (3H, s), 3.64 (3H, s). MS: m/z348.0 (M+H⁺).

Example V-102,5-Dimethoxy-N-(1-methyl-1H-pyrazol-3-yl)-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.09 (1H, s), 7.43 (1H, d), 7.23 (1H, t),7.14 (2H, d), 5.79 (1H, d), 3.81 (3H, s), 3.72 (3H, s), 3.63 (3H, s).MS: m/z 298.0 (M+H⁺).

Example V-115-Bromo-2-methoxy-N-(2-methyl-2H-pyrazol-3-yl)-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.27 (1H, s), 7.84 (1H, dd), 7.65 (1H, d),7.28 (1H, s), 7.26 (1H, t), 5.67 (1H, s), 3.94 (3H, s), 3.63 (3H, s).MS: m/z 346.0 (M+H⁺).

Example V-122,5-Dimethoxy-N-(2-methyl-2H-pyrazol-3-yl)-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.09 (1H, s), 7.25-7.21 (3H, m), 7.12 (1H,s), 5.66 (1H, d), 3.89 (3H, s), 3.72 (3H, s), 3.63 (3H, s). MS: m/z298.1 (M+H⁺).

Example V-135-Bromo-2-methoxy-N-(1-methyl-5-trifluoromethyl-1H-pyrazol-3-yl)-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.76 (1H, s), 7.78-7.82 (2H, m), 7.20 (1H,d), 6.45 (1H, s), 3.82 (3H, s), 3.79 (3H, s). MS: m/z 415.7 (M+H⁺).

Example V-142,5-Dimethoxy-N-(1-methyl-5-trifluoromethyl-1H-pyrazol-3-yl)-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.63 (1H, s), 7.27 (1H, d), 7.20-7.14 (2H,m), 6.41 (1H, s), 3.78 (3H, s), 3.76 (3H, s), 3.73 (3H, s). MS: m/z366.0 (M+H⁺).

Example V-15 5-Bromo-2-methoxy-N-thiazol-2-yl-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=12.75 (1H, s), 7.86 (1H, d), 7.72 (1H, dd),7.27 (1H, d), 7.14 (1H, d), 6.87 (1H, d), 3.70 (3H, s). MS: m/z 350.9(M+H⁺).

Example V-16 2,5-Dimethoxy-N-thiazol-2-yl-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=12.63 (1H, s), 7.33 (1H, d), 7.24 (1H, d),7.13-7.11 (2H, m), 6.84 (1H, d), 3.75 (3H, s), 3.64 (3H, s). MS: m/z301.0 (M+H⁺).

Example V-175-Bromo-2-methoxy-N-(5-methyl-[1,3,4]thiadiazol-2-yl)-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=13.95 (1H, s), 7.87 (1H, d), 7.78 (1H, dd),7.18 (1H, d), 3.74 (3H, s), 2.53 (3H, s). MS: m/z 365.9 (M+H⁺).

Example V-182,5-Dimethoxy-N-(5-methyl-[1,3,4]thiadiazol-2-yl)-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=13.82 (1H, s), 7.32 (1H, d), 7.14-7.12 (2H,m), 3.75 (3H, s), 3.65 (3H, s), 2.51 (3H, s). MS: m/z 316.0 (M+H⁺).

Example V-195-Bromo-2-methoxy-N-(5-trifluoromethyl-[1,3,4]thiadiazol-2-yl)-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.57 (1H, s), 7.87 (1H, d), 7.75 (1H, s),7.17 (1H, d), 3.69 (3H, s). MS: m/z 419.9 (M+H⁺).

Example V-20N-(5-Difluoromethyl-[1,3,4]thiadiazol-2-yl)-2,5-dimethoxy-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (CD₃OD, 400 MHz): δ=7.45 (1H, s), 7.00 (1H, t), 3.76 (3H, s),3.63 (3H, s). MS: m/z 370.0 (M+H⁺).

Example V-215-Bromo-2-methoxy-N-(5-methyl-[1,3,4]oxadiazol-2-yl)-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=13.48 (1H, s), 7.90 (1H, d), 7.75 (1H, dd),7.17 (1H, d), 3.76 (3H, s), 2.35 (3H, s). MS: m/z 349.9 (M+H⁺).

Example V-222,5-Dimethoxy-N-(5-methyl-[1,3,4]oxadiazol-2-yl)-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=13.28 (1H, s), 7.37 (1H, d), 7.14-7.11 (2H,m), 3.77 (3H, s), 3.70 (3H, s), 2.36 (3H, s). MS: m/z 300.0 (M+H⁺).

Example V-23 2,5-Dimethoxy-N-pyrazin-2-yl-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.30 (1H, s), 8.34 (1H, s), 8.21-8.17 (2H,m), 7.39 (1H, d), 7.19 (1H, dd), 7.11 (1H, d), 3.77 (3H, s), 3.68 (3H,s). MS: m/z 296.1 (M+H⁺).

Example V-24 5-Bromo-2-methoxy-N-pyridazin-3-yl-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=14.41 (1H, d), 8.35 (1H, d), 8.68 (1H, d),7.92 (1H, d), 7.77-7.71 (2H, m), 7.14 (1H, d), 3.66 (3H, s). MS: m/z346.0 (M+H⁺).

Example V-25 2,5-Dimethoxy-N-pyridazin-3-yl-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.00 (1H, s), 8.35 (1H, s), 8.15-8.12 (1H,m), 7.71 (1H, dd), 7.38 (1H, d), 7.13-7.08 (2H, m), 3.77 (3H, s), 3.60(3H, s). MS: m/z 296.1 (M+H⁺).

Example V-26 5-Bromo-2-methoxy-N-pyridazin-4-yl-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=14.37 (1H, s), 8.53 (1H, t), 8.31 (1H, s),7.89 (1H, d), 7.70 (1H, d), 7.38 (1H, dd), 7.12 (1H, d), 3.71 (3H, s).MS: m/z 345.9 (M+H⁺).

Example V-27 2,5-Dimethoxy-N-pyridazin-4-yl-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=14.25 (1H, s), 8.49-8.45 (1H, m), 8.27 (1H,s), 7.38-7.34 (2H, m), 7.11-7.07 (2H, m), 3.76 (3H, s), 3.65 (3H, s).MS: m/z 296.1 (M+H⁺).

Example V-28 5-Bromo-2-methoxy-N-pyrimidin-2-yl-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=14.36 (1H, s), 8.54 (1H, d), 8.33 (1H, s),7.89 (1H, d), 7.70 (1H, d), 7.37 (1H, t), 7.12 (1H, d), 3.71 (3H, s).MS: m/z 343.9 (M+H⁺).

Example V-29 2,5-Dimethoxy-N-pyrimidin-2-yl-benzenesulfonamide

This compound was prepared as described in Example V-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.69 (1H, s), 8.48-8.44 (2H, m), 7.43 (1H,d), 7.17 (1H, d), 7.11 (1H, d), 7.09 (1H, d), 7.02 (1H, d), 3.78 (3H,s), 3.76 (3H, s). MS: m/z 296.1 (M+H⁺).

Example VI Example VI-1 5-Bromo-2-methoxy-pyridine-3-sulfonic acidquinolin-3-ylamide

Step 1: To the mixture of pyridin-2-ylamine (10.0 g, 106 mmol) inacetone (200 mL), was added NBS (22.6 g, 127 mmol) portionwise at 0° C.The mixture was warmed to room temperature and stirred overnight.Solvent was evaporated in vacuum. The residue was purified by silica gelcolumn (DCM/MeOH, 20/1) to afford 18 g (yield: 98%) of5-bromo-pyridin-2-ylamine as yellow solid.

¹H NMR (DMSO-d6): δ=7.94 (1H, d), 7.61 (1H, dd), 6.43 (1H, d), 6.10 (2H,brs).

Step 2: The mixture of 5-bromo-pyridin-2-ylamine (8.0 g, 46.2 mmol) inClSO₃H (20 mL) was stirred at 200° C. for 4 h. After cooled to roomtemperature, the mixture was poured into ice water and neutralized withNaHCO₃ solid. The aqueous phase was extracted with EtOAc (50 mL×3). Theextracts were dried over Na₂SO₄ and concentrated in vacuum to give aresidue, which was purified by silica gel column (DCM/MeOH, 20/1) toafford 2.5 g (yield: 23%) of4-bromo-7-thia-2,8-diaza-bicyclo[4.2.0]octa-1,3,5-triene 7,7-dioxide.

¹H NMR (DMSO-d6): δ=9.11 (1H, brs), 8.47 (1H, d), 8.08 (1H, d).

Step 3: The mixture of4-bromo-7-thia-2,8-diaza-bicyclo[4.2.0]octa-1,3,5-triene 7,7-dioxide(1.0 g, 4.3 mmol), quinolin-3-ylamine (735 mg, 5.1 mmol) in pyridine (20mL) was stirred at room temperature overnight. Solvent was evaporated invacuum. The residue was washed with DCM (5 mL×2). The resulting solidwas collected by filtration to afford 800 mg (yield: 49%) of2-amino-5-bromo-pyridine-3-sulfonic acid quinolin-3-ylamide as whitesolid.

¹H NMR (DMSO-d6): δ=11.07 (1H, brs), 8.63 (1H, d), 8.23 (1H, d),7.98-7.93 (4H, m), 7.69-7.66 (1H, m), 7.60-7.58 (1H, m), 6.95 (2H, brs).MS: m/z 379.0 (M+H⁺).

Step 4: To the mixture of 2-amino-5-bromo-pyridine-3-sulfonic acidquinolin-3-ylamide (600 mg, 1.6 mmol) in concentrated HCl (40 mL) wasadded NaNO₂ (110 mg, 69 mmol) portionwise at 0° C. The mixture wasslowly warmed up to room temperature. The suspension was filtered toafford 1.0 g (75% purity on LCMS) of5-bromo-2-chloro-pyridine-3-sulfonic acid quinolin-3-ylamide as whitesolid. MS: m/z 397.9 (M+H⁺).

Step 5: The mixture of 5-bromo-2-chloro-pyridine-3-sulfonic acidquinolin-3-ylamide (300 mg crude), NaOMe (200 mg, 3.7 mmol) in MeOH (3mL) was stirred at 100° C. in a sealed tube for 2 h. The solvent wasevaporated in vacuum and the residue was purified by silica gel columnto afford 40 mg (two step yield: 21%) of5-bromo-2-methoxy-pyridine-3-sulfonic acid quinolin-3-ylamide as whitesolid.

¹H NMR (DMSO-d6): δ=11.01 (1H, brs), 8.69 (1H, d), 8.53 (1H, d), 8.33(1H, d), 8.00 (1H, d), 7.93-7.89 (2H, m), 7.66-7.63 (1H, m), 7.56-7.53(1H, m), 3.91 (3H, s). MS: m/z 393.6 (M+H⁺).

Example VI-22,5-Dimethoxy-N-methyl-N-(quinolin-3-yl)pyridine-3-sulfonamide

Step 1: To a solution of 5-bromo-2-methoxy-pyridine-3-sulfonic acidquinolin-3-ylamide (100 mg, 0.25 mmol) in DMF (5 mL) was added Pd(PPh₃)₄(10 mg), K₂CO₃ (70 mg, 0.5 mmol) and bis(pinacolato)diboron (127 mg, 0.5mmol), and the mixture was irradiated by microwave at 120° C. for 2 h.LCMS showed that the reaction was complete. The resultant was pouredinto water, and extracted with EtOAc (20 mL×3). The extracts were driedover Na₂SO₄ and concentrated in vacuum to give crude2-methoxy-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridine-3-sulfonicacid quinolin-3-ylamide.

Step 2: To crude2-methoxy-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-O-pyridine-3-sulfonicacid quinolin-3-ylamide in DCM (5 mL) was added H₂O₂ (2 mL) and AcOH(0.5 mL). The mixture was stirred at r.t for 1 h. TLC showed that thereaction was complete. The resultant was concentrated in vacuum andpurified directly by silica gel plates (PE/EtOAc, 1:1) to give 25 mg(two-step yield: 29%) of 5-hydroxy-2-methoxy-pyridine-3-sulfonic acidquinolin-3-ylamide as colorless solid.

¹H NMR (CD₃OD, 400 MHZ): δ=8.55 (1H, d), 7.92 (1H, d), 7.82 (1H, d),7.72-7.68 (2H, m), 7.57-7.53 (2H, m), 7.47-7.43 (1H, m), 3.82 (3H, s).MS: m/z 332 (M+H⁺).

Example VII Example VII-15-Bromo-2-methoxy-N-methyl-N-(quinolin-3-yl)benzenesulfonamide

To a solution of 5-bromo-2-methoxy-N-(quinolin-3-yl)benzenesulfonamide(100 mg, 0.26 mmol) in THF (2 mL) was added K₂CO₃ (70 mg, 0.52 mmol) andmethane iodide (74 mg, 0.52 mmol) at room temperature, then the mixturewas stirred at room temperature overnight. The solvent was removed invacuum. The residue was diluted with EtOAc (20 ml). The mixture waswashed with water, brine and dried over Na₂SO₄. The solution wasevaporated to dryness and purified by prep-HPLC (PE/EtOAc, 10/1) toafford 15 mg (yield: 14%) of5-bromo-2-methoxy-N-methyl-N-(quinolin-3-yl)benzenesulfonamide as whitesolid.

¹H NMR (DMSO-d6, 400 MHz) δ: 8.82 (1H, d), 8.23 (1H, d), 8.01 (1H, d),7.97 (1H, d), 7.83 (1H, dd), 7.82-7.73 (2H, m), 7.65 (1H, t), 7.21 (1H,d), 3.57 (3H, s), 3.38 (3H, s). MS: m/z 406.9 (M+H⁺)

Example VII-25-Bromo-2-methoxy-N-ethyl-N-(quinolin-3-yl)benzenesulfonamide

To a solution of 5-bromo-2-methoxy-N-(quinolin-3-yl)benzenesulfonamide(100 mg, 0.26 mmol) in DMF (2 mL) was added K₂CO₃ (70 mg, 0.52 mmol) andethyl bromide (40 mg, 0.31 mmol) at room temperature, then the mixturewas stirred at 80° C. overnight. After cooled to room temperature, thesolvent was removed in vacuum. The residue was diluted with EtOAc (20ml). The mixture was washed with water, brine and dried over Na₂SO₄. Thesolution was evaporated to dryness and purified by silica gel column(PE/EtOAc, 20/1) to afford 15 mg (yield: 14%) of5-bromo-2-methoxy-N-ethyl-N-(quinolin-3-yl)benzenesulfonamide as whitesolid.

¹H NMR (DMSO-d6, 400 MHz) δ: 8.68 (1H, d), 8.25 (1H, d), 8.01 (2H, t),7.85-7.76 (2H, m), 7.67-7.61 (2H, m), 7.27 (1H, d), 3.87 (2H, q), 3.77(3H, s), 1.06 (3H, t). MS: m/z 420.9 (M+H⁺)

Example VII-35-Bromo-2-methoxy-N-propyl-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (DMSO-d6, 400 MHz) δ: 8.70 (1H, d), 8.27 (1H, d), 8.01 (2H, t),7.84-7.75 (2H, m), 7.67-7.63 (2H, m), 7.26 (1H, d), 3.83-3.75 (5H, m),1.44-1.35 (2H, m), 0.87 (3H, t). MS: m/z 434.9 (M+H⁺)

Example VII-45-Bromo-N-isopropyl-2-methoxy-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (DMSO-d6, 400 MHz) δ: 8.43 (1H, d), 8.18 (1H, d), 8.06 (2H, t),7.89-7.81 (2H, m), 7.67 (1H, t), 7.61 (1H, d), 7.34 (1H, d), 4.65-4.59(1H, m), 3.99 (3H, s), 1.09 (6H, d). MS: m/z 435.0 (M+H⁺)

Example VII-55-Bromo-N-butyl-2-methoxy-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (DMSO-d6, 400 MHz) δ: 8.70 (1H, d), 8.26 (1H, d), 8.01 (2H, t),7.85-7.78 (2H, m), 7.67-7.63 (2H, m), 7.26 (1H, d), 3.83 (2H, t), 3.77(3H, s), 1.37-1.28 (4H, m), 0.82 (3H, t). MS: m/z 449.0 (M+H⁺)

Example VII-65-Bromo-N-benzyl-2-methoxy-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (DMSO-d6, 400 MHz) δ: 8.55 (1H, d), 7.99 (1H, d), 7.92 (1H, s),7.91 (1H, s), 7.71-7.60 (3H, m), 7.52 (1H, t), 7.30-7.18 (5H, m), 6.94(1H, d), 5.05 (2H, s), 3.88 (3H, s). MS: m/z 483.0 (M+H⁺)

Example VII-75-Bromo-2-methoxy-N-(2-methoxyethyl)-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (DMSO-d6, 400 MHz) δ: 8.67 (1H, d), 8.22 (1H, d), 7.80 (2H, t),7.84-7.75 (2H, m), 7.66-7.60 (2H, m), 7.28 (1H, d), 4.02 (2H, t), 3.84(3H, s), 3.41 (2H, t), 3.13 (3H, s). MS: m/z 451.0 (M+H⁺)

Example VII-85-Bromo-2-methoxy-N-(3-methoxypropyl)-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (CDCl₃, 400 MHz) δ: 8.66 (1H, d), 8.12 (1H, d), 8.09 (1H, d),7.91 (1H, d), 7.81 (1H, d), 7.74 (1H, td), 7.61-7.55 (2H, m), 6.87 (1H,d), 3.96 (2H, t), 3.75 (3H, s), 3.44 (2H, t), 3.23 (3H, s), 1.85-1.77(2H, t). MS: m/z 465.0 (M+H⁺)

Example VII-95-Bromo-N-(2-(dimethylamino)ethyl)-2-methoxy-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (DMSO-d6, 400 MHz) δ: 8.63 (1H, d), 8.13 (1H, d), 7.92 (1H, d),7.81 (1H, d), 7.73-7.67 (1H, m), 7.66-7.59 (2H, m), 7.58-7.54 (1H, m),7.07 (1H, d), 3.93 (2H, t), 3.69 (3H, s), 2.41 (2H, t), 2.13 (6H, s).MS: m/z 464.0 (M+H⁺)

Example VII-10N-allyl-5-bromo-2-methoxy-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (DMSO-d6, 400 MHz) δ: 8.68 (1H, d), 8.23 (1H, d), 7.99 (2H, t),7.84 (1H, dd), 7.81-7.75 (1H, m), 7.68-7.60 (2H, m), 7.28 (1H, d),5.83-5.74 (1H, m), 5.21-5.16 (1H, m), 5.07 (1H, d), 4.50 (2H, d), 3.81(3H, s). MS: m/z 433.0 (M+H⁺)

Example VII-115-Bromo-2-methoxy-N-(prop-2-yn-1-yl)-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (CDCl₃, 400 MHz) δ: 8.71 (1H, d), 8.16 (1H, d), 8.08 (1H, d),7.91 (1H, d), 7.82 (1H, d), 7.75 (1H, td), 7.65-7.57 (2H, m), 6.91 (1H,d), 4.71 (2H, d), 3.82 (3H, s), 2.24 (1H, t). MS: m/z 431.0 (M+H⁺)

Example VII-122,5-Dimethoxy-N-methyl-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (DMSO-d6, 400 MHz) δ: 8.79 (1H, d), 8.07-8.04 (2H, m), 7.78 (1H,d), 7.69 (1H, t), 7.55 (1H, t), 7.38 (1H, d), 7.04 (1H, dd), 6.89 (1H,d), 3.73 (3H, s), 3.63 (3H, s), 3.47 (3H, s). MS: m/z 359.1 (M+H⁺)

Example VII-13 2,5-Dimethoxy-N-ethyl-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (DMSO-d6, 400 MHz) δ: 8.67 (1H, d), 8.22 (1H, d), 8.00 (2H, t),7.78 (1H, td), 7.66-7.61 (1H, m), 7.23-7.20 (2H, m), 7.10 (1H, t), 3.87(2H, q), 3.70 (3H, s), 3.66 (3H, s), 1.06 (3H, t). MS: m/z 373.1 (M+H⁺).

Example VII-142,5-Dimethoxy-N-isopropyl-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (DMSO-d6, 400 MHz) δ: 8.42 (1H, d), 8.16 (1H, d), 8.05 (2H, t),7.83 (1H, td), 7.68-7.63 (1H, m), 7.32-7.23 (2H, m), 7.08 (1H, d),4.64-4.56 (1H, m), 3.92 (3H, s), 3.66 (3H, s), 1.09 (6H, d). MS: m/z387.1 (M+H⁺).

Example VII-15 2,5-Dimethoxy-N-butyl-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (DMSO-d6, 400 MHz) δ: 8.69 (1H, d), 8.23 (1H, d), 8.02-7.97 (2H,m), 7.80-7.74 (1H, m), 7.63 (1H, t), 7.22-7.20 (2H, m), 7.08 (1H, s),3.83 (2H, t), 3.72 (3H, s), 3.65 (3H, s), 1.40-1.26 (4 h, m), 0.81 (3H,t). MS: m/z 401.1 (M+H⁺)

Example VII-162,5-Dimethoxy-N-benzyl-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (CDCl₃, 400 MHz) δ: 8.57 (1H, d), 7.98 (1H, d), 7.90 (1H, d),7.69-7.63 (2H, m), 7.52-7.47 (1H, m), 7.32-7.18 (6H, m), 7.06 (1H, dd),7.00 (1H, d), 5.07 (2H, s), 3.88 (3H, s), 3.68 (3H, s). MS: m/z 435.1(M+H⁺)

Example VII-172,5-Dimethoxy-N-(2-methoxyethyl)-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (CDCl₃, 400 MHz) δ: 8.66 (1H, d), 8.11 (1H, d), 8.05 (1H, d),7.78 (1H, d), 7.71 (1H, td), 7.58-7.52 (1H, m), 7.25 (1H, d), 7.02 (1H,dd), 6.95 (1H, d), 4.08 (2H, t), 3.84 (3H, s), 3.66 (3H, s), 3.56 (2H,t), 3.27 (3H, s). MS: m/z 403.1 (M+H⁺)

Example VII-182,5-Dimethoxy-N-(3-methoxypropyl)-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (DMSO-d6, 400 MHz) δ: 8.71 (1H, d), 8.25 (1H, d), 8.01 (2H, t),7.82-7.76 (1H, m), 7.64 (1H, t), 7.25-7.22 (2H, m), 7.11 (1H, s), 3.90(2H, t), 3.73 (3H, s), 3.66 (3H, s), 3.35 (2H, t), 3.14 (3H, s),1.68-1.61 (2H, m). MS: m/z 417.1 (M+H⁺)

Example VII-19N-(2-(Dimethylamino)ethyl)-2,5-dimethoxy-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (DMSO-d6, 400 MHz) δ: 8.71 (1H, d), 8.24 (1H, d), 7.99 (1H, d),7.97 (1H, d), 7.77 (1H, t), 7.63 (1H, t), 7.25-7.20 (2H, m), 7.05 (1H,d), 4.16-4.00 (2H, m), 3.76 (3H, s), 3.63 (3H, s), 2.80-2.60 (2H, m),2.36 (6H, s). MS: m/z 416.2 (M+H⁺)

Example VII-20 N-Allyl-2,5-dimethoxy-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (CDCl₃, 400 MHz) δ: 8.61 (1H, d), 8.07-8.02 (2H, m), 7.78 (1H,d), 7.71 (1H, td), 7.55 (1H, t), 7.29 (1H, d), 7.04 (1H, dd), 6.96 (1H,d), 5.91-5.81 (1H, m), 5.15-5.06 (2H, m), 4.51 (2H, d), 3.82 (3H, s),3.68 (3H, s). MS: m/z 385.1 (M+H⁺)

Example VII-212,5-Dimethoxy-N-(prop-2-yn-1-yl)-N-(quinolin-3-yl)benzenesulfonamide

This compound was prepared as described in Example VII-2.

¹H NMR (CDCl₃, 400 MHz) δ: 8.70 (1H, d), 8.15 (1H, d), 8.07 (1H, d),7.80 (1H, d), 7.73 (1H, td), 7.57 (1H, t), 7.28 (1H, d), 7.07 (1H, dd),6.97 (1H, d), 4.73 (2H, d), 3.84 (3H, s), 3.69 (3H, s), 2.22 (1H, t).MS: m/z 383.1 (M+H⁺)

Example VIII Example VIII-15-Bromo-2-methoxy-N-(6-morpholinopyridin-3-yl)benzenesulfonamide

Step 1: To a solution of 2-chloro-5-nitropyridine (200 mg, 1.27 mmol) inDCM (10 mL) was added morpholine (115 mg, 1.27 mmol) and TEA (256 mg,2.54 mmol), and the mixture was stirred at r.t overnight. TLC showedthat the reaction was complete. The resultant was washed with water (100mL) and brine (100 mL), concentrated in vacuum and purified by silicagel column (PE/EtOAc, 10/1) to give 210 mg (yield: 79%) of4-(5-nitropyridin-2-yl)morpholine as a yellow solid. MS: m/z 210.1(M+H⁺).

Step 2: To a solution of 4-(5-nitropyridin-2-yl)morpholine (210 mg, 1mmol) in methanol (15 mL) was added Pd/C (20 mg), and the mixture washydrogenated at r.t under atmosphere pressure for 3 h. TLC showed thatthe reaction was complete. The resultant was filtered to remove Pd/C,and the filtrate was purified by silica gel column (PE/EtOAc, 1/1) togive 150 mg (yield: 84%) of 6-morpholinopyridin-3-amine as a brownsolid.

Step 3: To a solution of 6-morpholinopyridin-3-amine (80 mg, 0.45 mmol)in pyridine (5 mL) was added 5-bromo-2-methoxybenzene-1-sulfonylchloride (126 mg, 0.45 mmol) and DMAP (10 mg), and the mixture wasstirred at 60° C. overnight. LCMS showed that the reaction was complete.The resultant was concentrated in vacuum to remove pyridine and theresidue was diluted with DCM (20 mL). The mixture was washed with 1N HCl(15 mL), dried over Na₂SO₄ and concentrated in vacuum. The crude productwas purified by prep-TLC (DCM/MeOH, 15/1) to give 30 mg (yield: 16%) of5-bromo-2-methoxy-N-(6-morpholinopyridin-3-yl)benzenesulfonamide as awhite solid.

¹H NMR (DMSO-d6, 400 MHz): δ=9.72 (1H, brs), 7.80-7.76 (2H, m), 7.64(1H, d), 7.23 (1H, d), 7.20 (1H, d), 6.72 (1H, d), 3.94 (3H, s), 3.64(4H, t), 3.35-3.30 (4H, m). MS: m/z 428.0 (M+H⁺).

Example VIII-22,5-Dimethoxy-N-(6-morpholinopyridin-3-yl)benzenesulfonamide

¹H NMR (DMSO-d6, 400 MHz): δ=9.55 (1H, brs), 7.79 (1H, d), 7.25 (1H, d),7.17-7.12 (2H, m), 7.10 (1H, d), 6.70 (1H, d), 3.87 (3H, s), 3.68 (3H,s), 3.63 (4H, t), 3.33-3.29 (4H, m). MS: m/z 380.1 (M+H⁺).

Example VIII-3 5-Bromo-2-methoxy-N-(6-Amino-pyridin-3-yl)-benzenesulfonamide

Step 1: To a solution of 5-chloro-2-nitropyridine (3 g, 19 mmol) in EtOH(30 mL) was added saturated NH₃.H₂O (20 mL), the mixture was stirredunder 50 psi at 150° C. overnight. TLC showed that the reaction wascomplete. After the reaction mixture was cooled to r.t, the resultingsolid was collected by filtration. The solid was washed with PE (100 mL)to give 0.7 g (yield: 26%) of 6-nitro-pyridin-3-ylamine as a yellowsolid. MS: m/z 140.1 (M+H⁺).

Step 2: To a solution of 6-nitro-pyridin-3-ylamine (50 mg, 0.33 mmol) inDMF (3 mL) was added NaH (60% in mineral oil, 25 mg, 0.66 mmol), and themixture was stirred at r.t for 30 min. Then5-bromo-2-methoxy-benzenesulfonyl chloride (86 mg, 0.3 mmol) was addedand the mixture was stirred at 50° C. overnight. TLC showed that thereaction was complete. The resultant was concentrated in vacuum toremove DMF. The residue was diluted with DCM (30 mL) and the mixture waswashed with 1N HCl (30 mL×3). The organic layer dried over Na₂SO₄ andconcentrated to dryness in vacuum. The residue was purified by silicagel column (PE/EtOAc, 1/1) to give 30 mg (yield: 26%) of5-bromo-2-methoxy-N-(6-nitro-pyridin-3-yl)-benzenesulfonamide as a brownsolid.

Step 3: To a solution of5-bromo-2-methoxy-N-(6-nitro-pyridin-3-yl)-benzenesulfonamide (200 mg,0.5 mmol) in methanol (20 mL) was added SnCl₂.H₂O (450 mg, 2 mmol), andthe mixture was stirred at reflux for 24 h. TLC showed that the reactionwas complete. The resultant was concentrated in vacuum to remove MeOH.It was basified with saturated NaHCO₃ (10 mL). The suspension wasfiltered, and the filtrate was purified by pre-HPLC to give 56 mg(yield: 31%) of5-bromo-2-methoxy-N-(6-Amino-pyridin-3-yl)-benzenesulfonamide as a brownsolid.

¹H NMR (DMSO-d6): δ=9.46 (1H, brs), 7.74 (1H, dd), 7.60 (1H, d), 7.52(1H, d), 7.20 (1H, d), 7.06 (1H, dd), 6.29 (1H, d), 5.85 (2H, brs), 3.93(3H, s). MS: m/z 358.0 (M+H⁺).

Example VIII-4 2,5-Dimethoxy-N-(6-Amino-pyridin-3-yl)-benzenesulfonamide

This compound was prepared as described in Example VIII-3.

¹H NMR (CD₃OD-d6, 400 MHz): δ=7.58 (1H, dd), 7.48 (1H, d), 7.18 (1H, d),7.09-7.04 (2H, m), 6.79 (1H, d), 3.81 (3H, s), 3.65 (3H, s). MS: m/z310.1 (M+H⁺).

Example VIII-55-Bromo-2-methoxy-N-(6-methylamino-pyridin-3-yl)-benzenesulfonamide

¹H NMR (DMSO-d6): δ=9.47 (1H, brs), 7.79-7.75 (1H, m), 7.62-7.59 (2H,m), 7.22 (1H, d), 7.09 (1H, dd), 6.48 (1H, d), 6.30 (1H, d), 3.96 (3H,s), 2.67 (3H, d). MS: m/z 372.0 (M+H⁺).

Example VIII-62,5-Dimethoxy-N-(6-methylamino-pyridin-3-yl)-benzenesulfonamide

¹H NMR (DMSO-d6): δ=9.31 (1H, brs), 7.65 (1H, d), 7.21-7.17 (2H, m),7.14-7.10 (2H, m), 6.46-6.44 (1H, d), 6.33-6.31 (1H, d), 3.93 (3H, s),3.72 (3H, s), 2.69 (3H, d). MS: m/z 324.1 (M+H⁺)

Example VIII-75-Bromo-N-(6-(dimethylamino)pyridin-3-yl)-2-methoxybenzenesulfonamide

¹H NMR (DMSO-d6, 400 MHz): δ=9.56 (1H, brs), 7.77 (1H, dd), 7.72 (1H,d), 7.61 (1H, s), 7.22-7.17 (2H, m), 6.50 (1H, d), 3.95 (3H, s), 2.93(6H, s). MS: m/z 386.0 (M+H⁺)

Example VIII-8N-(6-(Dimethylamino)pyridin-3-yl)-2,5-dimethoxybenzenesulfonamide

¹H NMR (DMSO-d6, 400 MHz): δ=9.37 (1H, brs), 7.72 (1H, d), 7.21-7.14(3H, m), 7.07 (1H, d), 6.49 (1H, d), 3.89 (3H, s), 3.68 (3H, s), 2.91(6H, s). MS: m/z 338.1 (M+H⁺).

Example VIII-95-Bromo-N-(6-(diethylamino)pyridin-3-yl)-2-methoxybenzenesulfonamide

¹H NMR (DMSO-d6, 400 MHz): δ=9.51 (1H, brs), 7.77 (1H, dd), 7.68 (1H,d), 7.63 (1H, d), 7.23 (1H, d), 7.15 (1H, dd), 6.46 (1H, d), 3.96 (3H,s), 3.38 (4H, q), 1.03 (6H, t). MS: m/z 414.1 (M+H⁺).

Example VIII-10N-(6-(Diethylamino)pyridin-3-yl)-2,5-dimethoxybenzenesulfonamide

¹H NMR (DMSO-d6, 400 MHz): δ=9.37 (1H, brs), 7.75 (1H, d), 7.23-7.18(3H, m), 7.15-7.14 (1H, d), 6.51-6.48 (1H, d), 3.94 (3H, s), 3.75 (3H,s), 3.43 (4H, q), 1.08 (6H, t). MS: m/z 366.1 (M+H⁺).

Example VIII-115-Bromo-2-methoxy-N-(6-((2-methoxyethyl)amino)pyridin-3-yl)benzenesulfonamide

¹H NMR (DMSO-d6, 400 MHz): δ=9.48 (1H, brs), 7.76 (1H, dd), 7.63-7.57(2H, m), 7.22 (1H, d), 7.06 (1H, dd), 6.58 (1H, t), 6.38 (1H, d), 3.95(3H, s), 3.40-3.29 (4H, m), 3.23 (3H, s). MS: m/z 416.0 (M+H⁺).

Example VIII-122,5-Dimethoxy-N-(6-((2-methoxyethyl)amino)pyridin-3-yl)benzenesulfonamide

¹H NMR (DMSO-d6, 400 MHz): δ=9.29 (1H, brs), 7.59 (1H, d), 7.18-7.14(2H, m), 7.08-7.06 (2H, m), 6.54 (1H, t), 6.37 (1H, d), 3.89 (3H, s),3.69 (3H, s), 3.39-3.28 (4H, m), 3.23 (3H, s). MS: m/z 368.1 (M+H⁺).

Example VIII-135-Bromo-N-(6-((2-(dimethylamino)ethyl)amino)pyridin-3-yl)-2-methoxybenzenesulfonamide

¹H NMR (DMSO-d6, 400 MHz): δ=9.74 (1H, brs), 9.70 (1H, brs), 7.77 (1H,dd), 7.68 (1H, d), 7.65 (1H, d), 7.25-7.21 (2H, m), 6.52 (1H, d), 3.94(3H, s), 3.53 (2H, t), 3.20 (2H, t), 2.79 (6H, s). MS: m/z 429.1 (M+H⁺)

Example VIII-14N-(6-((2-(Dimethylamino)ethyl)amino)pyridin-3-yl)-2,5-dimethoxybenzenesulfonamide

¹H NMR (CD₃OD, 400 MHZ): δ=6.83 (1H, s), 6.34-6.20 (4H, m), 5.53 (1H,d), 3.06 (3H, s), 2.81 (3H, s), 2.59 (2H, t), 2.08 (2H, t), 1.75 (6H,s). MS: m/z 381.2 (M+H⁺).

Example VIII-155-Bromo-2-methoxy-N-(6-(phenylamino)pyridin-3-yl)benzenesulfonamide

¹H NMR (DMSO-d6, 400 MHz): δ=9.76 (1H, brs), 9.00 (1H, brs), 9.81-9.73(2H, m), 7.67 (1H, d), 7.56 (2H, d), 7.29 (1H, dd), 7.25-7.19 (3H, m),6.85 (1H, d), 6.72 (1H, d), 3.94 (3H, s). MS: m/z 434.0 (M+H+).

Example VIII-162,5-Dimethoxy-N-(6-morpholinopyridin-3-yl)benzenesulfonamide

¹H NMR (CDCl₃, 400 MHz): δ =7.77 (1H, d), 7.40 (1H, dd), 7.33-7.23 (5H,m), 7.06-6.97 (3H, m), 6.84 (1H, s), 6.73 (1H, d), 6.46 (1H, brs), 4.03(3H, s), 3.73 (3H, s). MS: m/z 386.1 (M+H⁺).

Example VIII-175-Bromo-2-methoxy-N-(3,4,5,6-tetrahydro-2H-[1,2]bipyridinyl-5′-yl)-benzenesulfonamide

¹H NMR (DMSO-d6, 400 MHz): δ=9.60 (1H, brs), 7.78-7.71 (2H, m), 7.63(1H, d), 7.22-7.16 (2H, m), 6.68 (1H, d), 3.93 (3H, s), 3.39 (4H, m),1.56-1.46 (6H, m). MS: m/z 426.0 (M+H⁺).

Example VIII-182,5-Dimethoxy-N-(3,4,5,6-tetrahydro-2H-[1,2′]bipyridinyl-5′-yl)-benzenesulfonamide

¹H NMR (DMSO-d6, 400 MHz): δ=9.44 (1H, brs), 7.73 (1H, d), 7.22-7.07(4H, m), 6.67 (1H, d), 3.88 (3H, s), 3.74 (3H, s), 3.41-3.32 (4H, m),1.55-1.45 (6H, m). MS: m/z 378.1 (M+H⁺).

Example VIII-195-Bromo-2-methoxy-N-[6-(4-methyl-piperazin-1-yl)-pyridin-3-yl]-benzenesulfonamide

¹H NMR (DMSO-d6, 400 MHz): δ=9.84 (1H, brs), 9.83 (1H, brs, TFA salt),7.82 (1H, d), 7.78 (1H, dd), 7.66 (1H, d), 7.32 (1H, dd), 7.21 (1H, d),6.84 (1H, d), 4.30-4.24 (2H, m), 3.93 (3H, s), 3.49-3.42 (2H, m),3.06-2.95 (4H, m), 2.81 (3H, s). MS: m/z 441.1 (M+H⁺).

Example VIII-202,5-Dimethoxy-N-[6-(4-methyl-piperazin-1-yl)-pyridin-3-yl]-benzenesulfonamide

¹H NMR (DMSO-d6, 400 MHz): δ=9.75 (1H, brs), 9.67 (1H, brs, TFA salt),7.84 (1H, d), 7.33 (1H, dd), 7.16-7.11 (3H, m), 6.83 (1H, d), 4.25-4.20(2H, m), 3.86 (3H, s), 3.69 (3H, s), 3.48-3.40 (2H, m), 3.09-2.95 (4H,m), 2.81 (3H, s). MS: m/z 393.2 (M+H⁺).

Example IX Example IX-15-Bromo-N-(6-hydroxyquinolin-3-yl)-2-methoxybenzenesulfonamide

Step 1: In a 2 L three-necked round-bottomed flask, equipped with athermometer, a dropping funnel, a mechanical stirrer and a gas vent, areplaced sodium nitrite (258 g, 3.74 mol) and water (250 mL). The contentsof the flask are heated and stirred to dissolve the solid. A solution ofmucobromic acid (258 g, 1 mol) in warm 95% ethanol (250 mL) is placed inthe dropping funnel and added dropwise with constant stirring over aperiod of 70-80 minutes. A mildly exothermic reaction occurs; thesolution in the flask becomes deep red, and gas is evolved. During theaddition, the temperature is kept at 54±1° C. by intermittentapplication of an ice bath to the flask. The mixture is stirred for anadditional 10 minutes at 54±1° C. While being stirred continuously, itis then cooled to 0-5° by application of an ice bath. The fine, yellowprecipitate is collected on a previously chilled Buchner funnel. Theslightly moist cake of crude product is transferred to a 1 L flask andheated to boiling with a mixture of 95% ethanol (400 mL) and water (100mL). The hot solution is filtered to remove a fine yellow solid, and theclear red filtrate is cooled to 0-5°. The recrystallized product iscollected by filtration and dried in air at room temperature to afford57 g (yield: 36%) of sodium nitromalonaldehyde monohydrate2-nitromalonaldehyde as tan needles.

Step 2: To a mechanically stirred solution of p-anisidine (74.5 g, 0.6mol) in 1.7 M aq.HCl (544 mL) was added a solution of2-nitromalonaldehyde (monohydrate, 63.0 g, 0.4 mol) in water (520 mL). Ayellow precipitate formed instantaneously and water was added tofacilitate stirring. After 10 minutes, the precipitate was filtered,washed with water and air-dried. The filter cake (98.0 g) was dried toconstant weight over P₂O₅ to give 68.0 g of enamine as a yellowamorphous solid. The enamine (68.0 g, 0.3 mol) was added to a vigorouslystirred suspention of p-anisidine hydrochloride (97.6 g, 0.6 mol) inacetic acid (612 mL) and the mixture was heated to refluxed under N₂.After 20 minutes thiophenol (6.73 g, 0.06 mol) was added and the mixtureheated at refluxed for 50 h. LC-MS showed the starting material wasmostly consumed. The reaction mixture was concentrated in vacuum, andneutralized with sat. NaHCO₃ to pH=9. The aqueous phase was extractedwith CHCl₃ (200 mL×3). The extracts were dried over Na₂SO₄ andconcentrated to dryness in vacuum. The crude was purified by silica gelcolumn (DCM/MeOH, 10/1) to give 30 g (yield: 25%) of6-methoxy-3-nitroquinoline as brown solid.

Step 3: A vigorously suspension of 6-methoxy-3-nitroquinoline (21.2 g,0.1 mol) in conc. HCl (320 mL) was heated to 50° C., the heating bathwas removed, and SnCl₂.H₂O (71.0 g, 0.3 mol) was added portion-wise over3 minutes. The mixture was stirred vigorously for an additional 10minutes and diluted with water to 1.0 L. The pH was brought to 9 byaddition of 5 M NaOH. The aqueous layer was cooled and extracted withEtOAc (150 mL×3). The extracts were washed brine and dried over Na₂SO₄.The mixture was concentrated in vacuum and the crude was purified bysilica gel column (DCM/MeOH, 10/1) to give 15 g (yield: 86%) of6-methoxyquinolin-3-amine as brown solid. MS: m/z 175.1 (M+H⁺).

Step 4: To a vigorously suspension of 6-methoxyquinolin-3-amine (1.0 g,5.7 mol) in anhydrous CH₂Cl₂ (20 mL) at −60° C. was added BBr₃ (3.0 mL)portionwise over 20 minutes. The mixture was stirred vigorously for anadditional 12 h and diluted with water to 100 mL. The aqueous layer wasextracted with EtOAc (25 mL×3). The extracts were washed brine and driedover Na₂SO₄. The mixture was concentrated to dryness and the residue waspurified by silica gel column (DCM/MeOH, 10/1) to give 500 mg (yield:55%) of 3-aminoquinolin-6-ol as brown solid. MS: m/z 161.1 (M+H⁺).

Step 5: A solution of 3-aminoquinolin-6-ol (150 mg, 0.9 mmol) and5-bromo-2-methoxybenzene-1-sulfonyl chloride (143 mg, 0.5 mmol) inpyridine (5 mL) was heated to 40° C. for 12 h. The reaction mixture wasconcentrated in vacuum and the residue was purified by silica gel column(PE/EtOAc, 10/1) to give 50 mg (yield: 25%) of5-bromo-N-(6-hydroxyquinolin-3-yl)-2-methoxybenzenesulfonamide as whitesolid.

¹H NMR (DMSO-d6, 400 MHz): δ=10.59 (1H, brs), 10.06 (1H, brs), 8.42 (1H,s), 7.85 (1H, s), 7.78-7.73 (3H, m), 7.20-7.17 (2H, m), 7.03 (1H, s),3.85 (3H, s). MS: m/z 409.2 (M+H⁺).

Example IX-25-Bromo-2-methoxy-N-(6-methoxyquinolin-3-yl)benzenesulfonamide

This compound was prepared as described in step 5, Example IX-1.

¹H NMR (CDCl₃, 400 MHz): δ=8.24 (1H, d), 7.91 (1H, d), 7.86 (1H, d),7.80 (1H, d), 7.49 (1H, dd), 7.21-7.19 (1H, m), 6.95 (1H, d), 6.83 (1H,d), 3.98 (3H, s), 3.86 (3H, s). MS: m/z 423.0 (M+H⁺).

Example IX-3 N-(6-Hydroxyquinolin-3-yl)-2,5-dimethoxybenzenesulfonamide

This compound was prepared as described in step 5, Example IX-1.

¹H NMR (CDCl₃, 400 MHz): δ=10.46 (1H, brs), 10.03 (1H, brs), 8.43 (1H,s), 7.73-7.70 (2H, m), 7.31 (1H, s), 7.18-7.11 (m, 3H), 6.99 (1H, s),3.79 (3H, s), 3.71 (3H, s). MS: m/z 361.2 (M+H⁺).

Example IX-43-(5-Bromo-2-methoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidmethyl ester

Step 1: All flasks used in the reaction were heated under vacuum for 30minutes and purged with N₂ for 10 minutes.1,1,1-trifluoro-N-phenyl-N-(trifluoromethylsulfonyl)methanesulfonamiden(214 mg, 0.6 mmol) was added into a solution ofN-(6-hydroxyquinolin-3-yl)-2,5-dimethoxybenzenesulfonamide (122 mg, 0.3mmol), DIPEA (0.12 mL, 0.7 mmol) in anhydrous THF (3 mL) at 0° C. andthen the reaction was stirred at room temperature for 12 h. LC-MS showedthe starting material was consumed completely. The mixture wasconcentrated in vacuum and the residue was purified by silica gel column(PE/EtOAc, 10/1) to give 100 mg of (yield: 66%)3-(5-bromo-2-methoxyphenylsulfonamido)quinolin-6-yltrifluoromethanesulfonate as white solid. MS: m/z 541.1 (M+H⁺).

Step 2: A solution of3-(5-bromo-2-methoxyphenylsulfonamido)quinolin-6-yltrifluoromethanesulfonate (100 mg, 0.18 mmol), Pd(dppf)Cl₂ (20 mg, 0.02mmol), NEt₃ (0.1 mL, 0.7 mmol) in MeOH (1 mL) and DMF (1 mL) was purgedwith CO for 3 times. The mixture was heated to 90° C. under 50 psi COatmosphere for 17 h. The suspension was filted, and the filtrate wasconcentrated in vacuum. The residue was purified silica gel column togive 60 mg (yield: 72%) of3-(5-bromo-2-methoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidmethyl ester.

¹H NMR (CDCl₃, 400 MHz): δ=8.65 (1H, s), 8.44 (1H, s), 8.08 (1H, d),8.01 (1H, s), 7.90-7.88 (2H, m), 7.54 (1H, d), 6.97 (1H, d), 3.87 (3H,s), 3.78 (3H, s). MS: m/z 450.9 (M+H⁺).

Example IX-53-(5-Bromo-2-methoxy-benzenesulfonylamino)-quinoline-6-carboxylic acid

1.0 M LiOH (0.1 mL, 0.1 mmol) was added into a solution of3-(5-bromo-2-methoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidmethyl ester (20 mg, 0.044 mmol) in THF (1 mL) and H₂O (1 mL) at 0° C.and the reaction was stirred at room temperature for 12 h. LC-MS showedthe starting material was consumed completely. The mixture wasconcentrated in vacuum and the remaining solution was acidified to pH=3with 2N HCl. The mixture was extracted with EtOAc (10 mL×3) and theextracts were dried over Na₂SO₄. The solvent was removed and the crudewas purified by silica gel column (PE/EtOAc, 1/1) to give 10 mg (yield:53%) of3-(5-Bromo-2-methoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidas white powder.

¹H NMR (CD₃OD, 400 MHz): δ=8.66 (1H, s), 8.44 (1H, s), 8.10 (1H, d),8.02 (1H, s), 7.91-7.88 (2H, m), 7.55 (1H, d), 6.98 (1H, d), 3.79 (3H,s). MS: m/z 434.7 (M−H⁺).

Example IX-63-(5-Bromo-2-methoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidamide

A suspension of3-(5-bromo-2-methoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidmethyl ester (50 mg, 0.11 mmol) in NH₃.H₂O (2.5 mL) in a sealed tube washeated at 80° C. for 2 h. LC-MS showed the starting material wasconsumed completely. The solution was concentrated in vacuum, and theresidue was purified by prep-HPLC to afford 9.8 mg (yield: 20%) of3-(5-Bromo-2-methoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidamide as orange solid.

¹H NMR (DMSO-d6, 400 MHz): δ=10.83 (1H, brs), 8.74 (1H, d), 8.41 (1H,d), 8.11 (1H, d), 8.03 (1H, d), 7.98-7.89 (2H, m), 7.88 (1H, d),7.75-7.73 (1H, m), 7.52 (1H, brs), 7.15 (1H, d), 3.83 (3H, s). MS: m/z436.0 (M+H⁺).

Example IX-73-(5-Bromo-2-methoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidmethylamide

A solution of3-(5-bromo-2-methoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidmethyl ester (50 mg, 0.11 mmol) in MeNH₂/MeOH (2.5 mL) in sealed tubewas heated at 80° C. for 2 h. LC-MS showed the starting material wasconsumed completely. The solution was concentrated in vacuum, and theresidue was purified via prep-HPLC to afford 14.4 mg (yield: 29%) of3-(5-bromo-2-methoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidmethylamide as white solid.

¹H NMR (DMSO-d6, 400 MHz): δ=10.81 (1H, brs), 8.76 (1H, d), 8.62 (1H,brs), 8.38 (1H, d), 8.04-8.02 (2H, m), 8.02 (1H, d), 7.89 (1H d),7.77-7.74 (1H, m), 7.16 (1H, d), 3.93 (3H, s), 2.83 (3H, d). MS: m/z450.0 (M+H⁺).

Example IX-83-(2,5-Dimethoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidmethyl ester

This compound was prepared as described in Example IX-4.

¹H NMR (CDCl₃, 400 MHz): δ=8.73 (1H, s), 8.51 (1H, s), 8.22 (1H, d),8.13 (1H, s), 8.06 (1H, d), 7.34 (1H, s), 7.36 (1H, s), 7.01-6.97 (2H,m), 4.03 (3H, s), 3.99 (3H, s), 3.72 (3H, s). MS: m/z 402.9 (M+H⁺).

Example IX-93-(2,5-Dimethoxy-benzenesulfonylamino)-quinoline-6-carboxylic acid

This compound was prepared as described in Example IX-5.

¹H NMR (CD₃OD, 400 MHz): δ=8.65 (1H, s), 8.39 (1H, s), 8.09 (1H, d),7.99 (1H, d), 7.87 (1H, d), 7.32 (1H, d), 7.18-7.11 (2H, m), 3.76 (3H,s), 3.64 (3H, s). MS: m/z 386.8 (M−H⁺).

Example IX-103-(2,5-Dimethoxy-benzenesulfonylamino)-quinoline-6-carboxylic acid amide

This compound was prepared as described in Example IX-6.

¹H NMR (DMSO-d6, 400 MHz): δ=10.67 (1H, brs), 8.77 (1H, d), 8.40 (1H,s), 8.12 (1H, s), 8.09-8.05 (1H, m), 8.03-7.93 (2H, m), 7.51 (1H, s),7.34 (1H, d), 7.16-7.10 (2H, m), 3.79 (3H, s), 3.72 (3H, s). MS: m/z388.1 (M+H⁺).

Example IX-113-(2,5-Dimethoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidmethylamide

This compound was prepared as described in Example IX-7.

¹H NMR (DMSO-d6, 400 MHz): δ=8.73 (1H, s), 8.58 (1H, s), 8.31 (1H, s),7.99-7.92 (3H, m), 7.35-7.34 (1H, d), 7.12-7.08 (2H, m), 3.76 (3H, s),3.47 (3H, s), 2.93 (3H, s). MS: m/z 401.9 (M+H⁺).

Example IX-123-(2,5-Dimethoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidpropylamide

This compound was prepared as described in Example IX-7.

¹H NMR (DMSO-d6, 400 MHz): δ=10.68 (1H, brs), 8.76 (1H, d), 8.61 (1H,t), 8.36 (1H, s), 8.03-8.00 (2H, m), 7.94 (1H, d), 7.34 (1H, d),7.16-7.09 (2H, m), 4.02 (3H, s), 3.97 (3H, s), 3.28-3.23 (2H, m),1.59-1.52 (2H, m), 0.91 (3H, t). MS: m/z 430.1 (M+H⁺).

Example IX-133-(2,5-Dimethoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidcyclopropylamide

To a solution of3-(2,5-dimethoxy-benzenesulfonylamino)-quinoline-6-carboxylic acid (40mg, 0.1 mmol), cyclopropylamine (30 mg, 0.5 mmol), NEt₃ (0.05 mL, 0.3mmol) and catalytic amount of DMAP in DMF (1 mL) was added HATU (150 mg,0.4 mmol) at 0° C. and then the reaction was stirred at room temperaturefor 12 h. LC-MS showed the starting material was consumed completely.The mixture was concentrated and the residue was purified by silica gelcolumn (PE/EtOAc, 10/1) to give 10 mg (yield: 23%) of3-(2,5-Dimethoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidcyclopropylamide as white powder.

¹H NMR (CD₃OD, 400 MHz): δ=8.64 (1H, s), 8.16 (1H, s), 8.03 (1H, s),7.96-7.84 (2H, m), 7.31 (1H, s), 7.00-6.94 (2H, m), 3.75 (3H, s), 3.63(3H, s), 2.82-2.77 (1H, m), 0.76-0.71 (2H, m), 0.59-0.55 (2H, m). MS:m/z 428.1 (M+H⁺).

Example IX-143-(2,5-Dimethoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidcyclohexylamide

This compound was prepared as described in Example IX-13.

¹H NMR (CD₃OD, 400 MHz): δ=8.61 (1H, d), 8.14 (1H, d), 7.99 (1H, d),7.92-7.84 (2H, m), 7.30 (1H, d), 6.99-6.94 (2H, m), 3.80-3.76 (1H, m),3.76 (3H, s), 3.62 (3H, s), 1.89-1.87 (2H, m), 1.74-1.71 (2H, m),1.34-1.26 (6H, m). MS: m/z 470.1 (M+H⁺).

Example IX-153-(2,5-Dimethoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidcyclohexylamide

Step 1: To a solution of methyl3-(2,5-dimethoxyphenylsulfonamido)quinoline-6-carboxylate (50 mg, 0.1mmol) in anhydrous THF (2 mL) was added LiAlH₄ (20 mg, 0.5 mmol) at 0°C., and the reaction was stirred at room temperature for 12 h. Thereaction was quenched with water (0.1 mL) and 15% NaOH (0.1 mL) at 0° C.The mixture was filtered through a pad of Na₂SO₄ and the filtrate wasconcentrated to dryness in vacuum. The residue was purified by silicagel column (PE/EtOAc, 10/1) to give 40 mg ofN-(6-(hydroxymethyl)quinolin-3-yl)-2,5-dimethoxybenzenesulfonamide(yield: 85%) as light yellow sticky liquid. MS: m/z 375.1 (M+H⁺).

Step 2: To a solution ofN-(6-(hydroxymethyl)quinolin-3-yl)-2,5-dimethoxybenzenesulfonamide (40mg, 0.1 mmol), NEt₃ (0.05 mL, 0.3 mmol) in THF (2 mL) at 0° C. was addedmethanesulfonyl chloride (50 mg, 0.4 mmol). The reaction was stirred atroom temperature for 12 h. LC-MS showed the starting material wasconsumed completely. The mixture was concentrated in vacuum and thecrude product was used directly for next step without furtherpurification.

Step 3: The crude product was dissolved in dimethylamine in THF (2.0 M).The reaction was stirred at room temperature overnight. LC-MS showed thestarting material was consumed completely. The mixture was concentratedin vacuum and the crude was purified by silica gel column (DCM/MeOH,10:1) to give 5 mg of3-(2,5-Dimethoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidcyclohexylamide (two-step yield: 12%) as white solid.

¹H NMR (CD₃OD, 400 MHz): δ=8.61 (1H, s), 7.99 (1H, d), 7.93 (1H, d),7.88 (1H, d), 7.60 (1H, dd), 7.31 (1H, d), 7.02-6.95 (2H, m), 4.35 (2H,s), 3.77 (3H, s), 3.64 (3H, s), 2.79 (6H, s). MS: m/z 402.0 (M+H⁺).

Example IX-16 3-(2,5-Dimethoxy-benzenesulfonylamino)-quinoline-6-carboxylic acidcyclohexylamide

This compound was prepared as described in Example IX-15.

¹H NMR (CD₃OD, 400 MHZ): δ=8.61 (1H, d), 8.02 (1H, d), 7.96-7.91 (2H,m), 7.84 (1H, d), 7.63 (1H, d), 7.54 (1H, dd), 6.98 (1H, d), 4.38 (2H,s), 3.81 (3H, s), 2.79 (6H, s). MS: m/z 451.0 (M+H⁺)

Example X Example X-1 5-Bromo-N-quinolin-3-yl-2-trifluoromethoxy-benzenesulfonamide

Step 1: A solution of 1-bromo-4-(trifluoromethoxy)benzene (1.0 g, 4.18mmol) in ClSO₃H (10 mL) was stirred at room temperature overnight. TLCshowed the starting material was consumed completely. The reactionmixture was poured into ice water (10 mL) and extracted with DCM (15×3).The combined organic layer was wash with brine (10 mL), dried overNa₂SO₄ and concentrated in vacuum to give 1 g (yield: 70%) of5-bromo-2-(trifluoromethoxy)benzene-1-sulfonyl chloride as colorlessoil.

Step 2: A mixture of 5-bromo-2-(trifluoromethoxy)benzene-1-sulfonylchloride (150 mg, 0.44 mmol), quinolin-3-ylamine (64 mg, 0.44 mmol) andDMAP (5 mg) in pyridine (5 mL) was stirred at 70° C. overnight. TLCshowed the starting material was consumed completely. The reactionmixture was diluted with EtOAc (20 mL), washed with water (20 mL), brine(10 mL) and dried over Na₂SO₄. The solution was concentrated in vacuumto give a residue, which was purified by prep-TLC (PE/EtOAc, 5/1) togive 33 mg (yield: 17%) of5-bromo-N-quinolin-3-yl-2-trifluoromethoxy-benzenesulfonamide asoff-white solid.

¹H NMR (CDCl₃, 400 MHz): δ=8.59 (1H, s), 8.07 (1H, s), 8.06-8.02 (2H,m), 7.78 (1H, d), 7.71-7.65 (2H, m), 7.60-7.55 (1H, m), 7.56-7.26 (1H,m), 7.05 (1H, brs). MS: m/z 447.0 (M+H⁺).

Example X-25-Fluoro-N-quinolin-3-yl-2-trifluoromethoxy-benzenesulfonamide

This compound was prepared as described in Example X-1.

¹H NMR (CDCl₃, 400 MHz): δ=8.58 (1H, s), 8.04-8.01 (2H, m), 7.78 (1H,d), 7.72-7.63 (2H, m), 7.56 (1H, t), 7.43-7.38 (1H, m), 7.32-7.25 (1H,m), 7.06 (1H, brs). MS: m/z 387.1 (M+H⁺).

Example X-35-Chloro-N-quinolin-3-yl-2-trifluoromethoxy-benzenesulfonamide

This compound was prepared as described in Example X-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.22 (1H, brs), 8.67 (1H, d), 8.04 (2H,dd), 7.94 (2H, d), 7.87-7.82 (1H, m), 7.68 (1H, t), 7.61-7.56 (2H, m).MS: m/z 403.0 (M+H⁺).

Example X-4 and X-55-Methyl-N-quinolin-3-yl-2-trifluoromethoxy-benzenesulfonamide and2-methyl-N-quinolin-3-yl-5-trifluoromethoxy-benzenesulfonamide

A mixture of 2-Methyl-5-trifluoromethoxy-benzenesulfonyl chloride and5-Methyl-2-trifluoromethoxy-benzenesulfonyl chloride (ratio: 1/1) wasobtained as described in Example X-1.

5-Methyl-N-quinolin-3-yl-2-trifluoromethoxy-benzenesulfonamide and2-methyl-N-quinolin-3-yl-5-trifluoromethoxy-benzenesulfonamide wereseparated by prep-HPLC.

For 5-methyl-N-quinolin-3-yl-2-trifluoromethoxy-benzenesulfonamide: ¹HNMR (CDCl₃, 400 MHz): δ=8.64 (1H, s), 8.04-8.00 (2H, m), 7.77-7.73 (2H,m), 7.65 (1H, t), 7.53 (1H, t), 7.34 (1H, dd), 7.29-7.24 (1H, m), 2.30(3H, s). MS: m/z 383.0 (M+H⁺).

For 2-methyl-N-quinolin-3-yl-5-trifluoromethoxy-benzenesulfonamide: ¹HNMR (CDCl₃, 400 MHz): δ=8.63 (1H, d), 8.02 (1H, d), 7.96 (1H, d), 7.90(1H, s), 7.70 (1H, d), 7.64 (1H, td), 7.54 (1H, t), 7.35-7.25 (2H, m),2.66 (3H, s). MS: m/z 383.0 (M+H⁺).

Example X-62-Methoxy-N-(quinolin-3-yl)-5-(trifluoromethoxy)benzenesulfonamide

¹H NMR (DMSO-d6, 400 MHz): δ=10.75 (1H, brs), 8.67 (1H, d), 8.00-7.84(3H, m), 7.77 (1H, d), 7.65-7.60 (2H, m), 7.55 (1H, t), 7.28 (1H, d),3.82 (3H, s). MS: m/z 399.1 (M+H⁺).

Example XI Example XI-15-chloro-N-[5-(4-methoxy-phenyl)-pyridin-3-yl]-2-trifluoromethoxy-benzenesulfonamide

Step 1: A solution of 1-chloro-4-(trifluoromethoxy)benzene (5.0 g, 25.0mmol) in ClSO₃H (35 mL) was stirred at room temperature overnight. TLCshowed the starting material was consumed completely. The reactionmixture was poured into ice water (50 mL) and extracted with DCM (50×3).The combined organic layer was wash with brine (50 mL), dried overNa₂SO₄ and concentrated in vacuum to give 3.9 g (yield: 53%) of5-chloro-2-(trifluoromethoxy)benzene-1-sulfonyl chloride as colorlessoil.

Step 2: A mixture of 5-bromopyridin-3-amine (300 mg, 1.74 mmol),4-methoxyphenylboronic acid (395 mg, 2.60 mmol), K₂CO₃ (240 mg, 5.10mmol) and Pd(PPh₃)₄ (197 mg, 0.17 mmol) in DMF/H₂O (5 mL/1 mL) waspurged with N₂ for 20 min. Then the mixture was stirred at 120° C. undermicrowave for 10 min. After cooled to room temperature, the solvent wasremoved in vacuum. The residue was diluted with EtOAc (30 mL). Themixture was washed with water, brine and dried over Na₂SO₄. The solutionwas evaporated to dryness and purified by silica gel column (DCM/MeOH,1/0-40/1) to afford 264 mg (yield: 44%) of5-(4-methoxyphenyl)pyridin-3-amine as white solid. MS: m/z 201.1 (M+H⁺).

Step 3: A mixture of 5-chloro-2-(trifluoromethoxy)benzene-1-sulfonylchloride (150 mg, 0.5 mmol), 5-(4-methoxyphenyl)pyridin-3-amine (100 mg,0.5 mmol) and DMAP (15 mg) in pyridine (5 mL) was stirred at 70° C. for4 h. TLC showed the starting material was consumed completely. Thereaction mixture was diluted with EtOAc (20 mL), washed with water (20mL), brine (10 mL) and dried over Na₂SO₄. The solution was concentratedin vacuum to give a residue, which was purified by prep-HPLC to give 6.5mg (yield: 3%)5-chloro-N-[5-(4-methoxy-phenyl)-pyridin-3-yl]-2-trifluoromethoxy-benzenesulfonamideas white solid.

¹H NMR (CD₃OD, 400 MHz): δ=8.48 (1H, s), 8.20 (1H, s), 8.05 (1H, s),7.77-7.75 (2H, m), 7.51-7.49 (3H, m), 7.04 (2H, d), 3.85 (3H, s). MS:m/z 458.9 (M+H⁺).

Example XI-22-Methoxy-N-(5-(4-methoxyphenyl)pyridin-3-yl)-5-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.62 (1H, brs), 8.49 (1H, s), 8.23 (1H,d), 7.34 (1H, d), 7.66-7.63 (2H, m), 7.49 (2H, d), 7.30 (1H, d), 7.04(2H, d), 3.88 (3H, s), 3.80 (3H, s). MS: m/z 454.8 (M+H⁺).

Example XI-35-Chloro-2-(trifluoromethoxy)-N-(5-(4-(trifluoromethoxy)phenyl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-D6, 400 MHz): δ=11.19 (1H, brs), 8.65 (1H, d), 8.32 (1H,d), 8.05 (1H, d), 7.86 (1H, dd), 7.77-7.72 (3H, m), 7.62 (1H, d), 7.50(2H, d). MS: m/z 512.7 (M+H⁺).

Example XI-45-Bromo-2-(trifluoromethoxy)-N-(5-(4-(trifluoromethoxy)phenyl)pyridin-3-yl)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.18 (1H, brs), 8.65 (1H, d), 8.31 (1H,d), 8.16 (1H, d), 8.00-7.97 (1H, m), 7.75-7.73 (3H, m), 7.54-7.49 (3H,m). MS: m/z 556.6 (M+H⁺).

Example XI-55-Chloro-N-(5-(3,4-dimethoxyphenyl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (CD₃OD, 400 MHz): δ=8.32 (1H, s), 8.18 (1H, d), 8.03 (1H, d),7.74-7.72 (2H, m), 7.52 (1H, d), 7.20 (1H, d), 6.67-6.64 (2H, m), 3.84(3H, s), 3.80 (3H, s). MS: m/z 488.9 (M+H⁺).

Example XI-65-Bromo-N-(5-(3,4-dimethoxyphenyl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.0 (1H, brs), 8.35 (1H, s), 8.21 (1H, d),8.10-8.09 (1H, m), 8.00-8.79 (1H, m), 7.58-7.55 (2H, m), 7.20-7.19 (1H,d), 6.68-6.63 (2H, m), 3.77 (3H, s), 3.73 (3H, s). MS: m/z 532.9 (M+H⁺).

Example XI-7N-(5-(3,4-Dimethoxyphenyl)pyridin-3-yl)-2-methoxy-5-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (CDCl₃, 400 MHz): δ =8.41 (1H, d), 8.02 (1H, d), 7.73 (1H, d),7.65-7.64 (1H, m), 7.30-7.29 (1H, m), 7.09-7.07 (1H, m), 6.98-6.96 (1H,d), 6.51-6.47 (2H, m), 3.98 (3H, s), 3.78 (3H, s), 3.70 (3H, s). MS: m/z484.9 (M+H⁺).

Example XI-85-Chloro-N-(5-(4-fluorophenyl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.11 (1H, brs), 8.61 (1H, d), 8.28 (1H,d), 8.05 (1H, d), 7.86 (1H, dd), 7.72 (1H, t), 7.68-7.58 (3H, m),7.36-7.31 (2H, m). MS: m/z 446.9 (M+H⁺).

Example XI-95-Bromo-N-(5-(4-fluorophenyl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.06 (1H, s), 8.61 (1H, s), 8.27 (1H, s),8.15 (1H, d), 8.00 (1H, d), 7.71-7.70 (1H, m), 7.66 (2H, dd), 7.54-7.51(1H, m), 7.35 (2H, t). MS: m/z 490.8 (M+H⁺).

Example XI-10N-(5-(4-Fluorophenyl)pyridin-3-yl)-2-methoxy-5-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.72 (1H, brs), 8.53 (1H, d), 8.29 (1H,d), 7.69 (1H, s), 7.68-7.58 (4H, m), 7.35-7.30 (3H, m), 3.89 (3H, s).MS: m/z 442.8 (M+H⁺).

Example XI-115-Bromo-N-(5-(4-cyanophenyl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.19 (1H, brs), 8.64 (1H, s), 8.32 (1H,d), 8.15 (1H, d), 7.98-7.94 (3H, m), 7.82-7.77 (3H, m), 7.51-7.49 (1H,m). MS: m/z 497.6 (M+H⁺).

Example XI-125-Chloro-N-(5-(furan-2-yl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (CD₃OD, 400 MHz): δ=8.53 (1H, s), 8.13 (1H, d), 8.03 (1H, d),7.83 (1H, t), 7.67 (1H, dd), 7.63 (1H, d), 7.47 (1H, d), 6.89 (1H, d),6.55 (1H, dd). MS: m/z 418.9 (M+H⁺).

Example XI-13N-(5-(Furan-2-yl)pyridin-3-yl)-2-methoxy-5-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.70 (1H, brs), 8.61 (1H, d), 8.20 (1H,d), 7.82 (1H, d), 7.75-7.70 (3H, m), 7.30 (1H, d), 7.04 (1H, d), 6.62(1H, dd), 3.87 (3H, s). MS: m/z 414.8 (M+H⁺).

Example XI-145-Chloro-N-(5-(thiophen-2-yl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (CD₃OD, 400 MHz): δ=8.42 (1H, d), 8.08 (1H, d), 7.95 (1H, d),7.68-7.67 (1H, m), 7.61 (1H, dd), 7.43-7.34 (3H, m), 7.07-7.05 (1H, m).MS: m/z 435.0 (M+H⁺).

Example XI-155-Bromo-N-(5-(thiophen-2-yl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.70 (1H, brs), 8.23 (1H, s), 8.06 (1H,d), 7.97 (1H, s), 7.75 (1H, d), 7.56 (1H, d), 7.46 (1H, s), 7.41 (1H,d), 7.36 (1H, d), 7.13 (1H, dd). MS: m/z 478.9 (M+H⁺).

Example XI-162-Methoxy-N-(5-(thiophen-2-yl)pyridin-3-yl)-5-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.68 (1H, brs), 8.59 (1H, d), 8.24 (1H,d), 7.75 (1H, d), 7.68-7.65 (3H, m), 7.52 (1H, dd), 7.32 (1H, d), 7.17(1H, dd), 3.89 (3H, s). MS: m/z 431.0 (M+H⁺).

Example XI-17N-([2,3′-Bipyridin]-5′-yl)-5-chloro-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.11 (1H, brs), 8.98 (1H, d), 8.71 (1H,d), 8.37 (1H, d), 8.21 (1H, dd), 8.03-7.83 (4H, m), 7.43 (1H, d), 7.41(1H, d). MS: m/z 430.0 (M+H⁺).

Example XI-18N-([2,3′-Bipyridin]-5′-yl)-5-bromo-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.12 (1H, brs), 8.95 (1H, s), 8.70 (1H,d), 8.35 (1H, d), 8.20 (1H, s), 8.13 (1H, d), 8.00-7.90 (3H, m), 7.50(1H, d), 7.44-7.40 (1H, m). MS: m/z 474.0 (M+H⁺).

Example XI-19N-([2,3′-Bipyridin]-5′-yl)-2-methoxy-5-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.71 (1H, brs), 8.89 (1H, d), 8.70-8.67(1H, m), 8.36 (1H, d), 8.20 (1H, t), 7.94-7.90 (2H, m), 7.73 (1H, d),7.63 (1H, dd), 7.44-7.40 (1H, m), 7.29 (1H, d), 3.87 (3H, s). MS: m/z426.0 (M+H⁺).

Example XI-205-Chloro-N-(5-(pyrazin-2-yl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.20 (1H, brs), 9.26 (1H, s), 8.94 (1H,s), 8.75 (1H, s), 8.67 (1H, s), 8.38-8.35 (1H, m), 8.18 (1H, s), 8.01(1H, s), 7.85-7.82 (1H, m), 7.58-7.55 (1H, m). MS: m/z 430.9 (M+H⁺).

Example XI-215-Bromo-N-(5-(pyrazin-2-yl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.25 (1H, brs), 9.30 (1H, d), 9.07 (1H,s), 8.78 (1H, d), 8.70 (1H, d), 8.43 (1H, d), 8.24 (1H, s), 8.15 (1H,d), 8.00-7.96 (1H, m), 7.52 (1H, d). MS: m/z 474.7 (M+H⁺).

Example XI-222-Methoxy-N-(5-(pyrazin-2-yl)pyridin-3-yl)-5-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.55 (1H, brs), 9.25 (1H, d), 8.99 (1H,d), 8.75 (1H, dd), 8.68 (1H, d), 8.43 (1H, d), 8.22 (1H, t), 7.74 (1H,d), 7.64 (1H, dd), 7.30 (1H, d), 3.89 (3H, s). MS: m/z 426.8 (M+H⁺).

Example XI-235-chloro-N-(5-(pyrimidin-2-yl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (CD₃OD, 400 MHz): δ=9.08 (1H, s), 8.79-8.75 (2H, m), 8.46 (1H,t), 8.28 (1H, d), 7.96 (1H, d), 7.57 (1H, dd), 7.34-7.30 (2H, m). MS:m/z 431.0 (M+H⁺).

Example XI-245-Bromo-N-(5-(pyrimidin-2-yl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

1H NMR (DMSO-d6, 400 MHz): δ=9.07 (1H, s), 8.94-8.90 (2H, m), 8.39-8.32(2H, m), 8.11 (1H, d), 7.90-7.85 (1H, m), 7.52-7.42 (2H, m). MS: m/z475.0 (M+H⁺).

Example XI-255-Chloro-N-(5-(thiazol-5-yl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-D6, 400 MHz): δ=11.19 (1H, brs), 9.25 (1H, d), 9.16 (1H,d), 8.34 (1H, d), 8.28 (1H, d), 8.11 (1H, t), 8.01 (1H, d), 7.86-7.84(1H, m), 7.61 (1H, d). MS: m/z 436.0 (M+H⁺).

Example XI-265-bromo-N-(5-(thiazol-5-yl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.54 (1H, brs), 9.24 (1H, d), 8.92 (1H,s), 8.36 (1H, s), 8.34 (1H, d), 8.14-8.10 (2H, m), 8.01 (1H, d), 7.52(1H, d). MS: m/z 479.8 (M+H⁺).

Example XI-272-Methoxy-N-(5-(thiazol-5-yl)pyridin-3-yl)-5-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.20 (1H, s), 9.23 (1H, d), 8.84 (1H, s),8.29-8.27 (2H, m), 8.15 (1H, s), 7.69 (1H, d), 7.63-7.62 (1H, m), 7.25(1H, d), 3.87 (3H, s). MS: m/z 431.7 (M+H⁺).

Example XI-285-Chloro-N-(5-(thiazol-4-yl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-D6, 400 MHz): δ=11.22 (1H, brs), 9.19 (1H, s), 8.68 (1H,s), 8.37 (1H, s), 8.25 (1H, s), 8.05 (1H, d), 7.86-7.84 (1H, m), 7.70(1H, s), 7.62-7.60 (1H, m). MS: m/z 435.5 (M+H⁺).

Example XI-295-Bromo-N-(5-(thiazol-4-yl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=11.22 (1H, brs), 9.19 (1H, s), 8.70 (1H,d), 8.38 (1H, d), 8.26 (1H, d), 8.16 (1H, d), 8.00 (1H, dd), 7.70 (1H,d), 7.53-7.50 (1H, m). MS: m/z 479.9 (M+H⁺).

Example XI-302-Methoxy-N-(5-(thiazol-4-yl)pyridin-3-yl)-5-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.72 (1H, brs), 9.18 (1H, s), 8.63 (1H,s), 8.33 (1H, s), 8.31 (1H, s), 7.66 (1H, d), 7.65-7.63 (2H, m), 7.33(1H, d), 3.87 (3H, s). MS: m/z 431.8 (M+H⁺).

Example XI-315-Bromo-N-(5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)-2-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (DMSO-d6, 400 MHz): δ=10.89 (1H, brs), 8.56 (1H, d), 8.20 (1H,s), 8.14 (1H, d), 8.08 (1H, d), 7.98 (1H, d), 7.85 (1H, s), 7.61 (1H,t), 7.53-7.51 (1H, m), 3.86 (3H, s). MS: m/z 477.0 (M+H⁺).

Example XI-322-Methoxy-N-(5-(1-methyl-1H-pyrazol-4-yl)pyridin-3-yl)-5-(trifluoromethoxy)benzenesulfonamide

This compound was prepared as described in Example XI-1.

¹H NMR (CDCl₃, 400 MHz): δ=8.41 (1H, d), 7.92 (1H, d), 7.66-7.63 (3H,m), 7.57 (1H, s), 7.30 (1H, dd), 7.07 (1H, s), 6.97 (1H, d), 3.98 (3H,s), 3.89 (3H, s). MS: m/z 429.0 (M+H⁺).

Example XII Example XII-1 Methyl5-(5-chloro-2-(trifluoromethoxy)phenylsulfonamido)nicotinate

¹H NMR (DMSO-d6, 400 MHz): δ=11.28 (1H, brs), 8.81 (1H, d), 8.57 (1H,dd), 8.02-7.98 (2H, m), 7.88 (1H, dd), 7.61 (1H, dd), 3.87 (3H, s). MS:m/z 411.0 (M+H⁺)

Example XII-2 5-(5-Bromo-2-(trifluoromethoxy)phenylsulfonamido)nicotinicacid

¹H NMR (DMSO-d6, 400 MHz): δ=13.58 (1H, brs), 11.24 (1H, brs), 8.79 (1H,s), 8.52 (1H, d), 8.01-7.83 (3H, m), 7.61 (1H, d). MS: m/z 397.0 (M+H⁺).

Example XII-3 Cyclohexyl5-(5-chloro-2-(trifluoromethoxy)phenylsulfonamido)nicotinate

¹H NMR (DMSO-d6, 400 MHz): δ=11.30 (1H, brs), 8.80 (1H, d), 8.55 (1H,d), 8.02-7.98 (2H, m), 7.90 (1H, dd), 7.62 (1H, dd), 4.96-4.94 (1H, m),1.86-1.82 (2H, m), 1.69-1.67 (2H, m), 1.56-1.37 (6H, m). MS: m/z 479.1(M+H⁺).

Example XII-4 Phenyl5-(5-chloro-2-(trifluoromethoxy)phenylsulfonamido)nicotinate

¹H NMR (DMSO-d6, 400 MHz): δ=11.39 (1H, brs), 8.98 (1H, d), 8.63 (1H,d), 8.15-8.13 (1H, m), 8.04 (1H, d), 7.90 (1H, dd), 7.64 (1H, dd),7.50-7.46 (2H, m), 7.35-7.28 (3H, m). MS: m/z 473.0 (M+H⁺).

Example XII-55-(5-Chloro-2-(trifluoromethoxy)phenylsulfonamido)nicotinamide

¹H NMR (DMSO-d6, 400 MHz): δ=11.14 (1H, brs), 8.75 (1H, s), 8.43-8.41(1H, m), 8.16 (1H, s), 7.98-7.81 (3H, m), 7.62-7.58 (2H, m). MS: m/z396.0 (M+H⁺).

Example XII-65-(5-Chloro-2-(trifluoromethoxy)phenylsulfonamido)-N-methylnicotinamide

¹H NMR (DMSO-d6, 400 MHz): δ=11.14 (1H, brs), 8.73 (1H, d), 8.67-8.64(1H, m), 8.43-8.42 (1H, m), 7.98 (1H, d), 7.89-7.82 (2H, m), 7.62 (1H,dd), 2.76 (3H, d). MS: m/z 410.0 (M+H⁺).

Example XII-75-(5-Chloro-2-(trifluoromethoxy)phenylsulfonamido)-N-propylnicotinamide

¹H NMR (DMSO-d6, 400 MHz): δ=11.14 (1H, brs), 8.73 (1H, s), 8.68-8.65(1H, m), 8.42 (1H, d), 7.99 (1H, d), 7.88-7.85 (2H, m), 7.62 (1H, dd),3.22-3.17 (2H, m), 1.54-1.48 (2H, m), 0.89-0.85 (3H, m). MS: m/z 438.0(M+H⁺).

Example XII-85-(5-Chloro-2-(trifluoromethoxy)phenylsulfonamido)-N-cyclohexylnicotinamide

¹H NMR (DMSO-d6, 400 MHz): δ=11.13 (1H, brs), 8.73 (1H, s), 8.44-8.41(2H, m), 7.99 (1H, d), 7.89-7.86 (2H, m), 7.62 (1H, dd), 3.74-3.70 (1H,m), 1.79-1.71 (4H, m), 1.34-1.24 (6H, m). MS: m/z 478.1 (M+H⁺).

Example XII-9 Methyl5-(5-bromo-2-(trifluoromethoxy)phenylsulfonamido)nicotinate

¹H NMR (DMSO-d6, 400 MHz): δ=11.28 (1H, brs), 8.80 (1H, d), 8.56-8.54(1H, m), 8.13 (1H, d), 8.02-7.99 (2H, m), 7.55-7.52 (1H, m), 3.87 (3H,s). MS: m/z 454.9 (M+H⁺).

Example XII-105-(5-Bromo-2-(trifluoromethoxy)phenylsulfonamido)nicotinic acid

¹H NMR (DMSO-d6, 400 MHz): δ=13.57 (1H, brs), 11.25 (1H, brs), 8.77 (1H,d), 8.51 (1H, d), 8.10 (1H, d), 8.01-7.97 (2H, m), 7.54 (1H, dd). MS:m/z 441.0 (M+H⁺).

Example XII-11 Cyclohexyl5-(5-bromo-2-(trifluoromethoxy)phenylsulfonamido)nicotinate

¹H NMR (DMSO-d6, 400 MHz): δ=11.28 (1H, brs), 8.80 (1H, d), 8.55 (1H,d), 8.13 (1H, d), 8.02-7.98 (2H, m), 7.55 (1H, dd), 4.97-4.94 (1H, m),1.86-1.82 (2H, m), 1.69-1.67 (2H, m), 1.57-1.37 (6H, m). MS: m/z 523.0(M+H⁺).

Example XII-12 Phenyl5-(5-bromo-2-(trifluoromethoxy)phenylsulfonamido)nicotinate

¹H NMR (DMSO-d6, 400 MHz): δ=11.38 (1H, brs), 8.98 (1H, d), 8.63 (1H,d), 8.17-8.13 (2H, m), 8.03 (1H, dd), 7.57 (1H, dd), 7.50-7.47 (2H, m),7.35-7.29 (3H, m). MS: m/z 517.0 (M+H⁺).

Example XII-135-(5-Bromo-2-(trifluoromethoxy)phenylsulfonamido)nicotinamide

¹H NMR (DMSO-d6, 400 MHz): δ=11.11 (1H, brs), 8.77 (1H, s), 8.43 (1H,d), 8.17 (1H, s), 8.10 (1H, d), 8.00 (1H, dd), 7.92-7.91 (1H, m), 7.64(1H, s), 7.54 (1H, dd). MS: m/z 440.0 (M+H⁺).

Example XII-145-(5-Bromo-2-(trifluoromethoxy)phenylsulfonamido)-N-methylnicotinamide

¹H NMR (DMSO-d6, 400 MHz): δ=11.14 (1H, brs), 8.73 (1H, d), 8.66 (1H,d), 8.42 (1H, d), 8.10 (1H, d), 8.01-7.98 (1H, m), 7.89-7.88 (1H, m),7.54 (1H, dd), 2.77 (3H, d). MS: m/z 454.0 (M+H⁺).

Example XII-15 Methyl5-(2-methoxy-5-(trifluoromethoxy)phenylsulfonamido)nicotinate

¹H NMR (DMSO-d6, 400 MHz): δ=10.89 (1H, brs), 8.73 (1H, d), 8.54 (1H,d), 7.99-7.98 (1H, m), 7.75-7.71 (2H, m), 7.30 (1H, d), 3.85 (3H, s),3.83 (3H, s). MS: m/z 407.0 (M+H⁺).

Example XII-16 Propyl5-(2-methoxy-5-(trifluoromethoxy)phenylsulfonamido)nicotinate

¹H NMR (DMSO-d6, 400 MHz): δ=10.89 (1H, brs), 8.73 (1H, d), 8.54 (1H,d), 8.00-7.99 (1H, m), 7.72-7.66 (2H, m), 7.32-7.29 (1H, d), 4.24-4.21(2H, m), 3.84 (3H, s), 1.72-1.67 (2H, m), 0.95 (3H, t). MS: m/z 434.0(M+H⁺).

Example XII-17 Cyclohexyl5-(2-methoxy-5-(trifluoromethoxy)phenylsulfonamido)nicotinate

¹H NMR (DMSO-d6, 400 MHz): δ=11.86 (1H, brs), 8.72 (1H, s), 8.53 (1H,d), 7.99 (1H, d), 7.72-7.65 (2H, m), 7.31 (1H, d), 4.97-4.91 (1H, m),3.86 (3H, s), 1.87-1.32 (10H, m). MS: m/z 475.1 (M+H⁺).

Example XII-18 Phenyl5-(2-methoxy-5-(trifluoromethoxy)phenylsulfonamido)nicotinate

¹H NMR (DMSO-d6, 400 MHz): δ=11.97 (1H, brs), 8.91 (1H, d), 8.61 (1H,d), 8.13-8.12 (1H, m), 7.76 (1H, d), 7.70-7.67 (1H, m), 7.50-7.46 (2H,m), 7.35-7.27 (4H, m), 3.86 (3H, s). MS: m/z 469.0 (M+H⁺).

Example XII-195-(2-Methoxy-5-(trifluoromethoxy)phenylsulfonamido)nicotinamide

¹H NMR (DMSO-d6, 400 MHz): δ=10.74 (1H, brs), 8.66 (1H, s), 8.40 (1H,d), 8.11 (1H, s), 7.89 (1H, s), 7.70 (1H, d), 7.64 (1H, dd), 7.57 (1H,s), 7.28 (1H, d), 3.83 (3H, s). MS: m/z 392.0 (M+H⁺).

Example XII-205-(2-Methoxy-5-(trifluoromethoxy)phenylsulfonamido)-N-methylnicotinamide

¹H NMR (DMSO-d6, 400 MHz): δ=10.71 (1H, brs), 8.63 (1H, s), 8.60 (1H,brs), 8.40 (1H, d), 7.88 (1H, t), 7.70-7.63 (2H, m), 7.29 (1H, d), 3.83(3H, s), 2.75 (3H, d). MS: m/z 406.0 (M+H⁺).

Example XII-215-(2-Methoxy-5-(trifluoromethoxy)phenylsulfonamido)-N-propylnicotinamide

¹H NMR (DMSO-d6, 400 MHz): δ=10.71 (1H, brs), 8.65 (1H, s), 8.64 (1H,brs), 8.40 (1H, d), 7.88 (1H, s), 7.70-7.64 (2H, m), 7.30 (1H, d), 3.84(3H, s), 3.21-3.16 (2H, m), 1.51-1.47 (2H, m), 0.86 (3H, t). MS: m/z434.0 (M+H⁺).

Example XII-225-(2-Methoxy-5-(trifluoromethoxy)phenylsulfonamido)-N-cyclohexylnicotinamide

¹H NMR (DMSO-d6, 400 MHz): δ=10.69 (1H, brs), 8.64 (1H, d), 8.41-8.36(2H, m), 7.86 (1H, s), 7.71-7.63 (2H, m), 7.30 (1H, d), 3.84 (3H, s),3.70-3.68 (1H, m), 1.82-1.56 (5H, m), 1.30-1.00 (5H, m). MS: m/z 474.1(M+H⁺).

Example XII-235-[(5-Methoxy-2-trifluoromethoxy-benzenesulfonyl)-methyl-amino]-nicotinicacid methyl ester

Step 1: 5-(5-Bromo-2-trifluoromethoxy-benzenesulfonylamino)-nicotinicacid methyl ester (200 mg, 0.44 mmol), bis(pinacolato)diboron (112 mg,0.44 mmol), Pd(dppf)Cl₂ (25 mg, 2.2% mmol), AcOK (86 mg, 0.88 mmol) werestirred in 1,4-dioxane (5 mL) at 100° C. under N₂ for 4 hours. Aftercooled to room temperature, the mixture was partitioned between EtOAc(20 mL) and water (10 mL). The aqueous phase was extracted with EtOAc(10 mL×3) and the combined organic phase was dried over Na₂SO₄. Thesolution was concentrated in vacuum to give 150 mg of mixture of boronicacid and boronic ester.

Step 2: The above mixture was dissolved in THF (5 mL) followed by NaOH(18 mg, 0.44 mmol), H₂O₂ (0.5 mL). The mixture was stirred at 50° C. for1 h. The solvent was concentrated in vacuum and the residue wasdissolved in water (10 mL). The mixture was extracted with EtOAc (10mL×3). The organic layer was washed with brine (10 mL) and dried overanhydrous Na₂SO₄. The solution was concentrated in vacuum. The residuewas purified by prep-TLC (PE/EtOAc, 3/1) to afford 60 mg (two-stepyield: 35%) of5-(5-hydroxy-2-trifluoromethoxy-benzenesulfonylamino)-nicotinic acidmethyl ester as white solid.

Step 3: To a solution of5-(5-hydroxy-2-trifluoromethoxy-benzenesulfonylamino)-nicotinic acidmethyl ester (60 mg, 0.15 mmol) in DMF (1 mL) was added K₂CO₃ (25 mg,0.18 mmol) and methyl iodide (42 mg, 0.30 mmol) at room temperature,then the mixture was stirred at 80° C. overnight. After cooled to roomtemperature, the solvent was removed in vacuum. The residue was dilutedwith EtOAc (20 ml). The mixture was washed with water, brine and driedover Na₂SO₄. The solution was evaporated to dryness and purified byprep-HPLC to afford 18 mg (yield: 29%) of5-[(5-methoxy-2-trifluoromethoxy-benzenesulfonyl)-methyl-amino]-nicotinicacid methyl ester as white solid.

¹H NMR (CD₃OD, 400 MHz): δ=8.88 (1H, d), 8.57 (1H, d), 8.09 (1H, t),7.33 (1H, d), 7.26 (1H, d), 7.20-7.17 (1H, m), 3.85 (3H, s), 3.74 (3H,s), 3.25 (3H, s). MS: m/z 421.1 (M+H⁺).

Example XIII Example XIII-15-Fluoro-2-methoxy-N-pyridin-3-yl-benzenesulfonamide

To a stirred ClSO₃H (50 mL) was added 1-fluoro-4-methoxy-benzene (10.0g, 79.4 mmol) dropwise at 25° C. The mixture was stirred at thistemperature for 3 hours. The mixture was poured into ice water (200 mL)and extracted with EtOAc (200 mL). The organic layer was washed withbrine (100 mL), dried over Na₂SO₄ and evaporated in vacuum to give 12 gof crude product. The crude was purified by silica gel columnchromatography (PE/EtOAc, 20/1) to give 10.0 g of5-fluoro-2-methoxy-benzenesulfonyl chloride (yield: 56%) as yellow oil.

The mixture of 5-fluoro-2-methoxy-benzenesulfonyl chloride (3 g, 13.4mmol), pyridine-3-ylamine (1.89 g, 13.3 mmol), TEA (2.7 g, 26.7 mmol) inanhydrous THF (30 mL) was stirred at room temperature overnight. To themixture was added water (50 mL). The mixture was extracted with EtOAc(50 mL×3). The organic layer was dried over Na₂SO₄. The solution wasconcentrated to dryness and the residue solid was re-crystallized fromDCM to give 2.3 g (yield: 52%) of5-fluoro-2-methoxy-N-pyridin-3-yl-benzenesulfonamide as yellow solid.

¹H NMR (DMSO-d₆, 400 MHz): δ=10.45 (1H, brs), 8.31 (1H, d), 8.22 (1H,dd), 7.55 (1H, dd), 7.47-7.51 (2H, m), 7.19-7.29 (2H, m), 3.85 (3H, s).MS: m/z 283.0 (M+H⁺).

Example XIII-2 5-Bromo-2-methoxy-N-pyridin-3-yl-benzenesulfonamide

To a stirred mixture of pyridine-3-ylamine (2.5 g, 8.8 mmol) and5-bromo-2-methoxy-benzenesulfonyl chloride (15.8 g, 55.6 mmol) inpyridine (60 mL) was added DMAP (187 mg, 0.88 mmol). The mixture wasstirred at 40° C. for 4 hours. The mixture was cooled, concentrated todryness in vacuum. The residue was diluted with MeOH (100 mL) andstirred for 30 minutes. The suspended solid was filtered and washed withmethanol (50 mL), evaporated in vacuum to give 12.1 g (yield: 70%) of5-(5-bromo-2-methoxy-benzenesulfonylamino)-nicotinic acid methyl esteras white solid.

¹H NMR (DMSO-d₆, 400 MHz): δ=10.43 (1H, brs), 8.30 (1H, d), 8.23 (1H,dd), 7.78-7.74 (2H, m), 7.48-7.51 (1H, m), 7.28 (1H, dd), 7.17 (1H, d),3.85 (3H, s). MS: m/z 342.8 (M+H⁺).

Example XIII-3 5-Chloro-2-methoxy-N-pyridin-3-yl-benzenesulfonamide

The procedure is similar to5-bromo-2-methoxy-N-pyridin-3-yl-benzenesulfonamide in Example XIII-2

¹H NMR (DMSO-d₆, 400 MHz): δ=10.44 (1H, brs), 8.30 (1H, d), 8.23 (1H,dd), 7.64-7.71 (2H, m), 7.48-7.51 (1H, m), 7.27 (1H, dd), 7.23 (1H, d),3.86 (3H, s). MS: m/z 298.9 (M+H⁺).

Example XIII-45-(5-Fluoro-2-trifluoromethoxy-benzenesulfonylamino)-nicotinamide

This compound was prepared as described in Example IV-16 and IV-17.

¹H NMR (DMSO-d₆, 400 MHz): δ=11.12 (1H, s), 8.76 (1H, d), 8.43 (1H, d),8.16 (1H, s), 7.92 (1H, s), 7.83 (1H, dd), 7.68-7.63 (3H, m). MS: m/z379.0 (M+H⁺).

Example XIII-5 5-(5-Fluoro-2-methoxy-benzenesulfonylamino)-nicotinamide

This compound was prepared as described in Example IV-16 and IV-17.

¹H NMR (DMSO-d₆, 400 MHz): δ=10.60 (1H, brs), 8.72 (1H, d), 8.44 (1H,d), 8.15 (1H, brs), 7.93 (1H, s), 7.64-7.61 (2H, m), 7.51-7.47 (1H, m),7.27-7.24 (1H, m), 3.84 (3H, s). MS: m/z 326.0 (M+H⁺).

Example XIII-65-Bromo-2-methoxy-N-[5-(2-methoxy-pyrimidin-5-yl)-pyridin-3-yl]-benzenesulfonamide

Step 1: The mixture of 5-bromo-pyriden-3-ylamine (2.6 g, 15 mmol),boronic acid (1.9 g, 12.5 mmol), Pd(PPh₃)₄ (1.45 g, 1.25 mmol), K₂CO₃(5.18 g, 37.5 mmol) in DMF (50 mL) was stirred at 120° C. under N₂overnight. The reaction mixture was concentrated in vacuum. The residuewas purified by chromatography on silica gel eluting with EA, to give1.2 g (yield: 47.5%) of 5-(2-methoxypyrimidn-5-yl)-3-pyridylamine asyellow solid.

¹H NMR (CDCl₃, 400 MHz): δ=8.70 (2H, s), 8.17 (1H, d), 8.13 (1H, d),7.07 (1H, dd), 4.07 (3H, s).

Step 2: To a stirred mixture of5-(2-methoxypyrimidn-5-yl)-3-pyridylamine (0.876 g, 4.34 mmol) and5-bromo-2-methoxy-benzenesulfonyl chloride (1.36 g, 4.77 mmol) inpyridine (10 mL) was added DMAP (26 mg, 0.22 mmol). The mixture wasstirred at 55° C. for 17 hours. The mixture was cooled, concentrated todryness. The residue was diluted with MeOH (100 mL) and stirred for 30minutes. The suspended solid was filtered and washed with methanol (5mL), evaporated in vacuum to dryness to give 0.96 g (yield: 49%) of5-(5-bromo-2-methoxy-benzenesulfonylamino)-nicotinic acid methyl esteras yellow solid.

¹H NMR (DMSO-d₆, 400 MHz): δ 10.36 (s, 1H), 8.80 (2H, m), 8.58 (1H, s),8.35 (1H, d), 7.88 (1H, d), 7.72-7.74 (2H, m), 7.17 (1H, d), 3.98 (3H,s), 3.83 (3H, s). MS: m/z 451 (M+H⁺).

Example XIII-75-Fluoro-2-methoxy-N-[5-(2-methoxy-pyrimidin-5-yl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example XIII-6

¹H NMR (DMSO-d₆, 400 MHz): δ 10.61 (1H, s), 8.85 (2H, s), 8.59 (1H, d),8.33 (1H, d), 7.75 (1H, dd), 7.66 (1H, dd), 7.49 (1H, td), 7.23 (1H,dd), 3.97 (3H, s), 3.85 (3H, s). MS: m/z 391 (M+H⁺).

Example XIII-85-Chloro-2-methoxy-N-[5-(2-methoxy-pyrimidin-5-yl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example XIII-6

¹H NMR (DMSO-d₆, 400 MHz): δ 10.6 (1H, brs), 8.85 (2H, m), 8.80 (1H, d),8.33 (1H, d), 7.79 (1H, d), 7.74 (1H, d), 7.67 (1H, dd), 7.24 (1H, d),3.97 (3H, s), 3.86 (3H, s). MS: m/z 407 (M+H⁺).

Example XIII-95-Chloro-2-methoxy-N-(6′-methoxy-[3,3]bipyridinyl-5-yl)-benzenesulfonamide

This compound was prepared as described in Example XIII-6

¹H NMR (DMSO-d₆, 400 MHz): δ=10.58 (1H, s), 8.55 (1H, d), 8.40 (1H, d),8.29 (1H, d), 7.93 (1H, dd), 7.78 (1H, d), 7.66 (1H, dd), 7.24 (1H, d),6.95 (1H, d), 3.90 (3H, s), 3.86 (3H, s).

Example XIII-105-Fluoro-2-methoxy-N-(6′-methoxy-[3,3]bipyridinyl-5-yl)-benzenesulfonamide

This compound was prepared as described in Example XIII-6

¹H NMR (DMSO-d₆, 400 MHz): δ=10.56 (1H, s), 8.54 (1H, d), 8.39 (1H, d),8.29 (1H, d), 7.93 (1H, dd), 7.62-7.68 (2H, m), 7.48-7.49 (1H, m), 7.23(1H, dd), 6.95 (1H, d), 3.90 (3H, s), 3.85 (3H, s). MS: m/z 390.1(M+H⁺).

Example XIII-115-Fluoro-2-methoxy-N-[5-(4-methoxy-phenyl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example XIII-6

¹H NMR (DMSO-d₆, 400 MHz): δ 10.51 (1H, s), 8.50 (1H, d), 8.24 (1H, d),7.66 (1H, t), 7.61 (1H, dd), 7.51 (2H, d), 7.48 (1H, dd), 7.22 (1H, dd),7.05 (2H, d), 3.85 (3H, s), 3.80 (3H, s). MS: m/z 389.1 (M+H⁺).

Example XIII-125-Chloro-2-methoxy-N-[5-(4-methoxy-phenyl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example XIII-6

¹H NMR (DMSO-d₆, 400 MHz): δ 10.52 (1H, s), 8.51 (1H, d), 8.24 (1H, d),7.77 (1H, d), 7.70-7.62 (2H, m), 7.51 (2H, d), 7.24 (1H, d), 7.06 (2H,d), 3.86 (3H, s), 3.80 (3H, s). MS: m/z 405 (M+H⁺).

Example XIII-135-Fluoro-2-methoxy-N-[5-(4-cyano-phenyl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example XIII-6

¹H NMR (DMSO-d₆, 400 MHz): δ 10.62 (1H, s), 8.62 (1H, d), 8.36 (1H, d),7.97 (2H, d), 7.88-7.71 (3H, m), 7.63 (1H, dd), 7.49 (1H, td), 7.23 (1H,dd), 3.83 (3H, s). MS: m/z 384 (M+H⁺).

Example XIII-145-Chloro-2-methoxy-N-[5-(4-cyano-phenyl)-pyridin-3-yl]-benzenesulfonamide

This compound was prepared as described in Example XIII-6

¹H NMR (DMSO-d₆, 400 MHz): δ 10.64 (1H, s), 8.62 (1H, d), 8.35 (1H, d),7.97 (2H, d), 7.82-7.75 (4H, m), 7.66 (1H, dd), 7.23 (1H, d), 3.84 (3H,s). MS: m/z 400.1 (M+H⁺).

BIOLOGICAL EXAMPLES Example I Assays for Agents that Inhibit TNAP

Compound Screening Library

A compound library was supplied by the NIH Molecular Libraries SmallMolecule Repository. Compounds were selected to represent diversifiedchemical space with clusters of closely related analogues to aid in theHTS-based structure-activity based relationship studies.

Expression and Preparation of Test Enzymes

Expression plasmids containing a secreted epitope-tagged TNAP weretransfected into COS-1 cells for transient expression. The medium wasreplaced with Opti-MEM 24 h later and serum-free media containingsecreted proteins was collected 60 hours after electroporation. Themedium was dialyzed against TBS containing 1 mM MgCl₂ and 20 mM ZnCl₂(to remove phosphate) and filtered through a 0.22 μm cellulose acetatefilter.

High Throughput Screening Assays

i. TNAP Colorimetric Assay

A TNAP stock solution was diluted 120-fold and about 12 μl of dilutedTNAP solution was dispensed into 96 well microtiter plates with halfarea bottom (Costar, Corning, N.Y.) by an auto dispenser (Matrix,Hudson, N.H.). A robotic liquid handler, Biomek™ FX (Beckman Coulter,Fullerton, Calif.) dispensed about 2.5 μl of each compound (dissolved in10% DMSO) from the library plates. Plates were incubated at roomtemperature for at least one hour to allow TNAP to interact with eachcompound prior to addition of about 10.5 μl substrate solution (1.19 mMpNPP). After about 30 minutes of incubation, A_(405 nm) was measuredwith a microtiter plate reader, Analyst™ HT (Molecular Devices,Sunnyvale, Calif.). Both the enzyme (TNAP) and substrate (pNPP) solutionwere made in diethanolamine (DEA) buffers; the final reaction contains 1M DEA-HCl buffer, pH about 9.8, containing about 1 mM MgCl₂ and about 20ZnCl₂. The concentration of TNAP and pNPP (final about 0.5 mM) wereadjusted to obtain A_(405 nm)˜0.4, while maintaining good sensitivity tothe known inhibitors levamisole and phosphate, used as positivecontrols. K_(m) obtained with a 1/120 dilution of TNAP and a fixedincubation period of about 30 minutes, was 0.58+0.081 mM.

ii. TNAP Luminescence Assay

Compound aliquots (4 μL @ 100 μM in 10% DMSO) were added with about 8 μLof TNAP working solution, prepared by 800-fold dilution of TNAP in2.5-fold assay buffer (250 mM DEA, pH 9.8, 2.5 mM MgCl₂, 0.05 mM ZnCl₂).CDP-star substrate solution (about 8 μl of 125 μM in water) was added toeach well. The final concentration of CDP-star was equal its K_(m) valuedetermined in the assay buffer. Plates (white 384-well small volumeGreiner 784075) were incubated at room temperature for about 0.5 hourand luminescence signal was measured using an En Vision plate reader(PerkinElmer). Levamisole (1 mM final concentration) or 2% DMSO wereutilized as positive and negative controls, respectively. Dose-responseconfirmation was performed under similar conditions using 10-point2-fold serial dilution of compounds.

Enzyme Kinetic Experiments—Phosphatase Selectivity Assay

To determine the inhibition selectivity for inhibitor candidates, humanTNAP, PLAP or TAP were added to microtiter plates followed by additionof the substrate pNPP (0.5 mM) and activity was measured in 1 M DEA-HClbuffer, pH 9.8 or in 1 M Tris-HCl buffer, pH 7.5, containing 1 mM MgCl₂and 20 μM ZnCl₂, in the presence of potential inhibitors (0-30 μM).TNAP, PLAP and IAP activities were adjusted to an approximateλA_(405 nm), equivalent to 1, measured after 30 min. Residual APactivity in the presence of inhibitors was expressed as percentage ofthe control activity. To investigate the mechanism of inhibition, doublereciprocal plots of enzyme activity (expressed as mA_(4O5 nm) min-1) vs.substrate concentration were constructed, in the presence of variousconcentrations of added inhibitors (0-30 μM). The y-axis intercepts ofthe 1/v vs. 1/[S] plots, were then plotted vs. [I] to graphicallyextract K_(i) values as the x-intercept in this plot. The numericalvalues from y- and x-intercepts were derived via linear regressionanalysis, using software Prism 3.02 (GraphPad Software, Calif.). Theseanalyses were performed, using pNPP as a substrate in 1 M DEA-HClbuffer, pH 9.8, as well as in 1 M Tris-HCl buffer, pH 7.5, to determineK_(i) at optimal and physiological pH respectively. Inhibitors werefurther tested and sorted based on their kinetic properties at pH 7.4using PPi, the relevant natural substrate of TNAP. In this part of thestudy, pyrophosphate sodium salt (99% ACS reagent, Sigma-Aldrich, StLouis, Mo.) was used as a substrate. Amounts of released phosphate weremeasured using the Biomol Green Reagent (Biomol Research Laboratories,Inc., Plymouth Meeting, Pa.). Finally, to document the potency ofselected inhibitors in physiological media, TNAP inhibition by compoundsof Formula I-IV (0-30 μM) was studied at pH 7.4, during catalysis of 0.1mM pNPP, in the presence of increasing concentrations of Na₂HPO₄ (0-10mM) and pyrophosphate (0-40 mM).

Compound docking was performed using the Flexx program, part of theSyby1 package from Trios, Inc. Formal charges were used for protein andcompound atoms. Heteroatoms (phosphate, zincs and magnesium) wereconsidered as part of the pocket while docking.

TNAP Activity in Plasma

Compounds of Formula I-IV described herein were added to Grenier1536-well clear plates. 1.5 μl of 4× buffer and substrate mixture wasadded by MultiDrop Combi to the plates. 4.5 μl of mouse or human plasmawas then added to the wells with a Bravo liquid handler. The 4× bufferand substrate mix consisted of 400 mM Tris, 80 μm ZnCl₂, 4 mM MgCl₂ andeither 4 mM paranitrophenol phosphate (pNPP) or 8 mM phenolphthaleinmonophosphate (PPMP) as substrate. The compounds and substrate wereincubated in the plates with human plasma for 6 to 30 hours at roomtemperature with the assay plate sealed. The duration of steady-statecatalysis depended on the phosphatase activity of the plasma. For pNPPsubstrate, OD₃₈₀ measurements were taken and the plasma catalytic ratewas calculated. For PPMP substrate, color developer consisting of sodiumcarbonate and sodium hydroxide was added to adjust pH such thatphenolphthalein color deepened but remained stable prior to OD₅₅₅measurements and plasma catalytic rate calculation. Using twosubstrates, compounds that optically interfere with thespectrophotometric assay were filtered out.

Table 1 below shows assay data for certain compounds of Formula I-IVdescribed herein.

For the TNAP (PPi) data, “A” indicates an IC50 of less than 0.1 μM, “B”indicates an IC50 of greater than or equal to 0.1 μM and less than orequal to 1 μM, and “C” indicates and IC50 of greater than 1 μM.

For the human plasma TNAP data (pNPP), “A” indicates an IC50 of lessthan 5 μM, “B” indicates an IC50 of greater than or equal to 5 μM andless than or equal to 50 μM, and “C” indicates an IC50 of greater than50 μM.

TABLE 1 Biological Activity of Compounds of Formula I-IV. TNAP TNAPhuman plasma (PPi) (pNPP) EXAMPLE IC50 (μM) IC50 (μM) Example I-1 A AExample I-2 B A Example I-3 A A Example I-4 B A Example I-5 C ExampleI-6 B Example I-7 B B Example I-8 B Example I-9 B Example I-10 C ExampleI-11 C Example I-12 C Example I-13 C Example I-14 B B Example I-15 CExample I-16 C Example I-17 C Example I-18 C Example I-19 C Example I-20C Example I-21 C Example I-22 C Example I-23 C Example II-1 A A ExampleII-2 A B Example II-3 A B Example II-4 C C Example II-5 A Example II-6 BC Example II-7 B Example II-8 C Example II-9 B Example II-10 A ExampleII-11 C Example II-12 C Example II-13 A B Example II-14 B Example II-15A A Example II-16 B Example II-17 B Example II-18 C Example II-19 CExample II-20 B Example II-21 B Example II-22 C Example II-23 B ExampleIII-1 A B Example III-2 B B Example III-3 A B Example III-4 B ExampleIII-5 B Example III-6 B Example III-7 A A Example III-8 A A ExampleIII-9 A A Example III-10 B C Example III-11 B C Example III-12 B CExample III-13 B C Example III-14 B C Example III-15 B C Example III-16A C Example III-17 A C Example III-18 A C Example III-19 B C ExampleIII-20 B C Example III-21 B C Example III-22 B C Example III-23 B BExample III-24 B C Example III-25 A Example III-26 A Example III-27 AExample III-28 A Example III-29 A C Example III-30 A B Example III-31 AB Example III-32 A Example III-33 A B Example III-34 B Example III-35 AExample III-36 A Example III-37 B B Example III-38 B C Example III-39 BC Example III-40 A Example III-41 A Example III-42 A Example III-43 BExample III-44 B Example III-45 B Example III-46 A Example III-47 AExample III-48 A Example III-49 B Example III-50 B Example III-51 BExample III-52 B Example III-53 B Example III-54 B Example III-55 AExample III-56 A Example III-57 A Example III-58 A Example III-59 AExample III-60 A Example III-61 B B Example III-62 A Example III-63 AExample III-64 A A Example III-65 A Example III-66 A Example III-67 AExample III-68 A Example III-69 A Example III-70 A B Example III-71 A BExample III-72 A B Example III-73 A Example III-74 A Example III-75 AExample III-76 A B Example III-77 A C Example III-78 A C Example III-79A Example III-80 A Example III-81 A Example III-82 A Example III-83 AExample III-84 A Example III-85 A Example III-86 A Example III-87 AExample III-88 A A Example III-89 A A Example III-90 A A Example III-91B Example III-92 A B Example III-93 A B Example III-94 A Example III-95A Example III-96 A Example IV-1 A A Example IV-2 A A Example IV-3 A BExample IV-4 A B Example IV-5 A Example IV-6 A Example IV-7 A A ExampleIV-8 A B Example IV-9 A Example IV-10 A Example IV-11 A A Example IV-12A Example IV-13 A Example IV-14 A Example IV-15 A Example IV-16 B AExample IV-17 A A Example IV-18 B A Example IV-19 B B Example IV-20 A BExample IV-21 B A Example IV-22 B Example IV-23 B C Example IV-24 C BExample IV-25 B B Example IV-26 A A Example IV-27 B Example IV-28 BExample IV-29 A Example IV-30 A Example IV-31 B Example IV-32 B ExampleIV-33 B Example IV-34 B Example IV-35 B Example IV-36 B Example IV-37 BExample IV-38 B Example IV-39 A Example IV-40 B Example IV-41 B ExampleIV-42 B Example IV-43 B B Example IV-44 A Example V-1 C Example V-2 CExample V-3 C C Example V-4 C Example V-5 B Example V-6 B Example V-7 CC Example V-8 C C Example V-9 C Example V-10 C Example V-11 C ExampleV-12 C Example V-13 C Example V-14 C Example V-15 C Example V-16 CExample V-17 C Example V-18 C Example V-19 B C Example V-20 C C ExampleV-21 C Example V-22 C Example V-23 C C Example V-24 B Example V-25 CExample V-26 C Example V-27 C Example V-28 C Example V-29 C Example VI-1A B Example VI-2 A B Example VII-1 C Example VII-2 C Example VII-3 CExample VII-4 C Example VII-5 C Example VII-6 C Example VII-7 C ExampleVII-8 C Example VII-9 C Example VII-10 C Example VII-11 C Example VII-12C Example VII-13 C Example VII-14 C Example VII-15 C Example VII-16 CExample VII-17 C Example VII-18 C Example VII-19 C Example VII-20 CExample VII-21 C Example VIII-1 C Example VIII-2 C Example VIII-3 C CExample VIII-4 C B Example VIII-5 B Example VIII-6 C Example VIII-7 CExample VIII-8 C Example VIII-9 B Example VIII-10 C Example VIII-11 CExample VIII-12 C Example VIII-13 C Example VIII-14 C Example VIII-15 CC Example VIII-16 C C Example VIII-17 C Example VIII-18 C ExampleVIII-19 C Example VIII-20 C Example IX-1 A Example IX-2 A Example IX-3 AExample IX-4 A C Example IX-5 B B Example IX-6 A B Example IX-7 A AExample IX-8 B C Example IX-9 C C Example IX-10 B B Example IX-11 B BExample IX-12 A B Example IX-13 B B Example IX-14 A A Example IX-15 B AExample IX-16 A A Example X-1 A Example X-2 B Example X-3 A C ExampleX-4 A Example X-5 C Example X-6 A B Example XI-1 A C Example XI-2 A CExample XI-3 B C Example XI-4 B C Example XI-5 B C Example XI-6 A CExample XI-7 A C Example XI-8 A C Example XI-9 B C Example XI-10 B CExample XI-11 B C Example XI-12 A B Example XI-13 A B Example XI-14 B CExample XI-15 B C Example XI-16 A C Example XI-17 B C Example XI-18 B CExample XI-19 B C Example XI-20 B C Example XI-21 B C Example XI-22 B CExample XI-23 C C Example XI-24 B C Example XI-25 B C Example XI-26 A CExample XI-27 B C Example XI-28 A B Example XI-29 A B Example XI-30 A BExample XI-31 A B Example XI-32 A B Example XII-1 B B Example XII-2 B BExample XII-3 A C Example XII-4 A B Example XII-5 B B Example XII-6 B BExample XII-7 C C Example XII-8 B C Example XII-9 B B Example XII-10 A BExample XII-11 A C Example XII-12 A B Example XII-13 A A Example XII-14B B Example XII-15 B B Example XII-16 A C Example XII-17 A C ExampleXII-18 A B Example XII-19 B A Example XII-20 B B Example XII-21 C BExample XII-22 B C Example XII-23 C C Example XIII-1 A Example XIII-2 AExample XIII-3 A Example XIII-4 A Example XIII-5 A Example XIII-6 AExample XIII-7 A Example XIII-8 A Example XIII-9 B Example XIII-10 BExample XIII-11 B Example XIII-12 B Example XIII-13 B Example XIII-14 B

A A

C

C A < 0.1 μM A < 5 μM 0.1 μM ≦ B ≦ 1 5 μM ≦ B ≦ 50 μM μM C > 1 μM C > 50μM

Example 2 Mouse Model of Medial Vascular Calcification

A conditional knock-in model of GACI was generated via a Cre-mediatedexpression of a TNAP knock-in transgene (FIG. 1) to assess whetheroverexpression of TNAP in vascular smooth muscle cells (VSMC) issufficient to cause medial vascular calcification (MVC). A vectorcontaining the human TNAP coding sequence under the control of theubiquitous CAG (CMV immediate early enhancer/chicken (3-actin promoterfusion) promoter was produced. This construct also included aloxP-flanked “stop cassette” between the promoter and transgene toprevent overexpression in the absence of Cre recombinase. Thistransgenic construct was introduced into the hypoxanthinephosphoribosyltransferase (Hprt) locus on the X chromosome. Hprt encodesa constitutively expressed housekeeping enzyme involved in nucleotidemetabolism and is located in a genomic region with an open chromatinstructure that allows permanent access to transcription factors,allowing it to be constitutively active. Targeted insertion at thislocus also overcomes any position effects that may occur with randomintegration methods. This mouse line, named Hprt^(ALPL), was used toexamine the effects of TNAP overexpression in VSMCs. Hprt^(ALPL/ALPL)female mice were bred with male mice expressing Cre-recombinase underthe control of the VSMC-specific transgelin promoter (Tagln-Cre)(Boucher P et al., Science 300: 329-322 (2003)). By breeding Tagln-Crehomozygous males with Hprt^(ALPL/ALPL) homozygous females, all maleoffspring are [Hprt^(ALPL/Y); Tagln-Cre^(+/−)] and all females areheterozygous for both transgenes.

Characterization of 30-day-old male [Hprt^(ALPL/Y); Tagln-Cre^(+/−)]mice shows that overexpression of TNAP in vascular smooth muscle cells(VSMCs) leads to severe calcification, as shown by Alizarin Red and vonKossa staining, as well as X-ray and μCT analysis of the aorta (FIG. 2).Calcification becomes visible by X-ray by 14 days of age andprogressively worsens with age until death before 3 months of age (FIG.3A). The male [Hprt^(ALPL/Y); Tagln-Cre^(+/−)] mice also showed 15-foldhigher serum alkaline phosphatase activity than WT mice. Necropsyrevealed greatly enlarged hearts in the [Hprt^(ALPL/Y); Tagln-Cre^(+/−)]mice, indicating cardiac insufficiency as the likely cause of death inthese animals (FIG. 3B).

Heterozygous female mice developed MVC more slowly and lived longer. Nopremature death or cardiac hypertrophy was observed in heterozygousfemale mice, up to 180 days of age. Alkaline phosphatase activity wasfound to be increased 4 fold in the heterozygous females as compared towild-type mice. The data suggests that upregulation of TNAP expressionin the vasculature is sufficient to cause MVC.

Example 3 Ex Vivo Study of Mineralization of Aortic Explants

Ex Vivo Studies—Aortas are carefully cleaned, and cut into two segmentsfor culture. Because of some heterogeneity within aortas, we routinelyuse eight aortic sections (four mice) per experimental variable. Thesmooth muscle cells remain viable in culture for at least two weeks withonly minor histological changes. Aortic explants are cultured inDulbecco's Modified Eagle Medium containing 60,000 cpm/mL of 45Ca and2.9 mM NaH₂PO₄, to induce mineralization within nine to twelve days.After this culture period, aortic rings are dehydrated and treated withHCl to liberate calcium, which is then measured by liquid scintillationcounting. In addition, we will culture VSMCs isolated from the aortas ofeach mouse model and measure PPi output and changes in gene expressionthat might be influenced by changes in either local or systemic ePPiconcentrations, as per published methods (50).

Example 4 In Vivo [Hprt_(ALPL); Tagln-Cre] Mouse Model

[Hprt_(ALPL/Y); Tagln-Cre_(+/−)] male or [Hprt_(ALPL/WT);Tagln-Cre_(+/−)] female mice and WT littermate controls are injectedwith a compound of Formula I-IV and its effectiveness in preventing MVCis assessed, while also any secondary effects of the treatment onskeletal mineralization or other organs is assessed. Evaluation of invivo efficacy is accomplished by assessing the drug metabolism andpharmacokinetic (DMPK) properties of the TNAP inhibitor of Formula I-IVand assessing PK/PD relationships.

[Hprt_(ALPL/Y); Tagln-Cre_(+/−)] male mice are dosed as a model of GACI.The treatment is initiated at 7 days-of-age and treatment continued for7 weeks until 60 days of age. Residual plasma TNAP activity is used as asurrogate biomarker for treatment efficacy (see FIG. 9, PreliminaryResults). Improved survival and changes in the degree of MVC and bonemineralization status is assessed by X-ray, CT, histomorphometry anddynamic histomorphometry and detailed histopathological examination ofvascular and skeletal tissues is performed. Furthermore, a detailedhistopathological examination of soft tissues is performed, withparticular focus on liver and kidney, two organs that express TNAP inhumans under physiological conditions. Renal function is studied bymeasuring serum potassium, sodium, blood urea nitrogen, and creatinine.Hepatic function is examined by measuring albumin, liver transaminases,bilirubin, and gamma glutamyl transpeptidase levels. Serum phosphate,calcium and PTH levels are assessed to ensure that the compound ofFormula I-IV does not change phosphate or calcium homeostasis. Serialechocardiography, measured by the Vevo 770 Micro-ultrasound System, PWVand blood pressure are used to monitor changes in cardiovascularfunction during treatment. Changes in concentrations of PPi in plasmaand urine are monitored, as well as other biochemical parameters.

[Hprt_(ALPL/WT); Tagln-Cre_(+/−)] female mice are used as a model ofadult MVC. Here, dosing is initiated at 30 days, in some instances at 14days-of-age (depending on when MVC becomes apparent) and the female miceare treated for four to eight weeks.

Example 5 Effect of Test Compound in Mouse Model of Medial VascularCalcification

[Hprt^(ALPL/Y); Tagln-Cre^(+/−)] male or [Hprt^(ALPL/WT);Tagln-Cre^(+/−)] female mice and wild-type littermate controls areinjected with a Test Compound of Formula I-IV in order to assess theeffectiveness of a Test Compound in preventing medial vascularcalcification. Characterization of levels of medial vascularcalcification is performed as described above, with Alizarin Red and vonKossa staining, as well as X-ray and μCT analysis of the aorta.Secondary effects on skeletal or other organ mineralization due to theTest Compound are assessed.

Example 6 Pharmacokinetic Data of a Compound of Formula I-IV in Mice

The bioavailability and plasma pharmacokinetic properties in mice of acompound of Formula I-IV is measured following intravenous,intraperitoneal, subcutaneous, intramuscular and oral administration.Three wild-type mice per time point per route of administration aredosed at various levels. At least one group is dosed intravenously andat least one group is dosed by an extravascular route in order to assessoral bioavailability. Blood is sampled from each group at frequentintervals (e.g. 0.25, 0.5, 1, 2, 4, 6, 8, 12, 24, and 48 hours) andplasma levels of the test compound are assayed using liquidchromatography coupled with tandem mass spectrometry (LC-MS/MS). Theplasma-concentration time data are analyzed to obtain thepharmacokinetic profile, including the area under the curve (AUC).Comparison of the AUCs after extravascular and intravenous dosing can beused to construct a time versus concentration graph from which we candetermine half-life of the compound, clearance, volume of distribution,total exposure, and maximal concentrations.

Example 7 Vascular Calcification Clinical Trial

Human Clinical Trial of the Safety and/or Efficacy of a TNAP inhibitor(e.g., a compound of Formula I-IV, or a pharmaceutically acceptable saltthereof) therapy.

Objective: To determine the safety, pharmacokinetics, and efficacy ofadministered TNAP inhibitor (e.g., a compound of Formula I-IV, or apharmaceutically acceptable salt thereof).

Study Design: This will be a Phase I, single-center, open-label,non-randomized dose escalation study followed by a Phase II study invascular calcification patients (for example uremic patients). Thediagnosis of vascular calcification is confirmed by a coronary arterycalcium score of greater than 50. Patients must not have received otherinvestigational agents within 3 months of study initiation. Fertilepatients must agree to use adequate contraception throughout the studyand for 18 months after cessation of treatment with a TNAP inhibitor(e.g., a compound of Formula I-IV). Patients must not be undertakingrenal replacement therapy. Patients must also not have had a recentfracture (within the last 3 months). In addition, patients must not havean abnormal rhythm of the heart. Patients must also not currently betaking osteoporosis medication. In addition, patients must not havehypocalcaemia or pres-existing dental diseases. All subjects areevaluated for safety and all blood collections for pharmacokineticanalysis are collected as scheduled. All studies are performed withinstitutional ethics committee approval and patient consent.

Phase I: Patients receive (e.g., intravenous, oral, ip, or the like)TNAP inhibitor (e.g., a compound of Formula I-IV, or a pharmaceuticallyacceptable salt thereof) daily for 4 weeks. Cohorts of 3-6 patientsreceive escalating doses of TNAP inhibitor (e.g., a compound of FormulaI-IV, or a pharmaceutically acceptable salt thereof). Escalation willnot be performed until all patients in the previous dose cohort havebeen treated for 4 weeks and until results obtained 4 weeks aftertreatment initiation do not reveal toxicity. Doses of TNAP inhibitor(e.g., a compound of Formula I-IV, or a pharmaceutically acceptable saltthereof) may be held or modified for toxicity based on assessments asoutlined below. Dose escalation is considered complete, if 2 patientsexperience a Grade 3 Adverse Event (AE) or if 1 patient experiencesGrade 4 AE at a particular cohort.

Phase II: Patients receive TNAP inhibitor (e.g., a compound of FormulaI-IV, or a pharmaceutically acceptable salt thereof) as in phase I at asuitable dose below the dose used in the final cohort. Treatmentcontinues throughout a 24-month study period during which clinical(which includes safety and tolerability) assessments are performed.

Blood Sampling: Serial blood is drawn by direct vein puncture before andafter administration TNAP inhibitor (e.g., a compound of Formula I-IV,or a pharmaceutically acceptable salt thereof). Venous blood samples (5mL) for determination of serum concentrations are obtained in-hospitalduring a 24-hour period. Each serum sample is divided into two aliquots.All serum samples are stored at −20° C. Serum samples are shipped on dryice.

Pharmacokinetics: Patients undergo plasma/serum sample collection forpharmacokinetic evaluation in-hospital during a 24-hour period.Pharmacokinetic parameters are calculated by model independent methodson a Digital Equipment Corporation VAX 8600 computer system using thelatest version of the BIOAVL software. The following pharmacokineticsparameters are determined: peak serum concentration (C_(max)); time topeak serum concentration (t_(max)); area under the concentration-timecurve (AUC) from time zero to the last blood sampling time (AUC₀₋₇₂)calculated with the use of the linear trapezoidal rule; and terminalelimination half-life (t_(1/2)), computed from the elimination rateconstant. The elimination rate constant is estimated by linearregression of consecutive data points in the terminal linear region ofthe log-linear concentration-time plot. The mean, standard deviation(SD), and coefficient of variation (CV) of the pharmacokineticparameters are calculated for each treatment. The ratio of the parametermeans (preserved formulation/non-preserved formulation) is calculated.

Patient Response: The primary outcome measure is safety andtolerability, based on conventional laboratory and clinical assessments.The secondary outcome measure is the assessment of changes arterialstiffness measured by pulse wave velocity, changes in vascularcalcification on CT scans of superficial femoral artery and aorta, andchanges in serum calcium and phosphate levels. Cardiovascular events,including myocardial ischemia, myocardial infarction, cardiac failure,stroke, and/or peripheral vascular disease are also assessed.

Example 8 Ankylosing Spondylitis Clinical Trial

Human Clinical Trial of the Safety and/or Efficacy of a TNAP inhibitor(e.g., a compound of Formula I-IV, or a pharmaceutically acceptable saltthereof) therapy.

Objective: To determine the safety, pharmacokinetics, and efficacy ofadministered TNAP inhibitor (e.g., a compound of Formula I-IV, or apharmaceutically acceptable salt thereof).

Study Design: This will be a Phase I, single-center, open-label,non-randomized dose escalation study followed by a Phase II study inankylosing spondylitis patients. Patients must have a diagnosis of ASaccording to the modified New York Criteria for ankylosing spondylitis,have had active AS based on the opinion of a physician for at leastthere months, and have active AS with a BASDAI>=4 at the time of thescreening visit. Patients must not have received other investigationalagents within 3 months of study initiation. Fertile patients must agreeto use adequate contraception throughout the study and for 18 monthsafter cessation of treatment with a TNAP inhibitor (e.g., a compound ofFormula I-IV). Patients must not have a history of or currentinflammatory joint disease of origin other than AS, e.g., rheumatoidarthritis, systemic lupus erythematosus, etc. In addition, patients mustnot have hypocalcaemia or pres-existing dental diseases. All subjectsare evaluated for safety and all blood collections for pharmacokineticanalysis are collected as scheduled. All studies are performed withinstitutional ethics committee approval and patient consent.

Phase I: Patients receive (e.g., intravenous, oral, ip, or the like)TNAP inhibitor (e.g., a compound of Formula I-IV, or a pharmaceuticallyacceptable salt thereof) daily for 4 weeks. Cohorts of 3-6 patientsreceive escalating doses of TNAP inhibitor (e.g., a compound of FormulaI-IV, or a pharmaceutically acceptable salt thereof). Escalation willnot be performed until all patients in the previous dose cohort havebeen treated for 4 weeks and until results obtained 4 weeks aftertreatment initiation do not reveal toxicity. Doses of TNAP inhibitor(e.g., a compound of Formula I-IV, or a pharmaceutically acceptable saltthereof) may be held or modified for toxicity based on assessments asoutlined below. Dose escalation is considered complete, if 2 patientsexperience a Grade 3 Adverse Event (AE) or if 1 patient experiencesGrade 4 AE at a particular cohort.

Phase II: Patients receive TNAP inhibitor (e.g., a compound of FormulaI-IV, or a pharmaceutically acceptable salt thereof) as in phase I at asuitable dose below the dose used in the final cohort. Treatmentcontinues throughout a 24-month study period during which clinical(which includes safety and tolerability) assessments are performed.

Blood Sampling: Serial blood is drawn by direct vein puncture before andafter administration TNAP inhibitor (e.g., a compound of Formula I-IV,or a pharmaceutically acceptable salt thereof). Venous blood samples (5mL) for determination of serum concentrations are obtained in-hospitalduring a 24-hour period. Each serum sample is divided into two aliquots.All serum samples are stored at −20° C. Serum samples are shipped on dryice.

Pharmacokinetics: Patients undergo plasma/serum sample collection forpharmacokinetic evaluation in-hospital during a 24-hour period.Pharmacokinetic parameters are calculated by model independent methodson a Digital Equipment Corporation VAX 8600 computer system using thelatest version of the BIOAVL software. The following pharmacokineticsparameters are determined: peak serum concentration (C_(max)); time topeak serum concentration (t_(max)); area under the concentration-timecurve (AUC) from time zero to the last blood sampling time (AUC₀₋₇₂)calculated with the use of the linear trapezoidal rule; and terminalelimination half-life (t_(1/2)), computed from the elimination rateconstant. The elimination rate constant is estimated by linearregression of consecutive data points in the terminal linear region ofthe log-linear concentration-time plot. The mean, standard deviation(SD), and coefficient of variation (CV) of the pharmacokineticparameters are calculated for each treatment. The ratio of the parametermeans (preserved formulation/non-preserved formulation) is calculated.

Patient Response: The primary outcome measure is safety andtolerability, based on conventional laboratory and clinical assessments.The secondary outcome measure is the assessment of change of theAnkylosing Spondylitis Disease Activity Score (ASDAS) and the BathAnkylosing Spondylitis Disease Activity Index (BASDAI).

Example 9 Pseudoxanthoma Elasticum Clinical Trial

Human Clinical Trial of the Safety and/or Efficacy of a TNAP inhibitor(e.g., a compound of Formula I-IV, or a pharmaceutically acceptable saltthereof) therapy.

Objective: To determine the safety, pharmacokinetics, and efficacy ofadministered TNAP inhibitor (e.g., a compound of Formula I-IV, or apharmaceutically acceptable salt thereof).

Study Design: This will be a Phase I, single-center, open-label,non-randomized dose escalation study followed by a Phase II study inpseudoxanthoma elasticum patients. The diagnosis of pseudoxanthomaelasticum must be confirmed by biopsy (documenting some calcification ofelastic fibers) and the patient must have a clinical disease severitygrade of at least “1” (poorly defined, barely visible macules) atscreening. Patients must not have received other investigational agentswithin 3 months of study initiation. Fertile patients must agree to useadequate contraception throughout the study and for 18 months aftercessation of treatment with a TNAP inhibitor (e.g., a compound ofFormula I-IV). Patients must not be undertaking renal replacementtherapy. Patients must also not have had a recent fracture (within thelast 3 months). In addition, patients must not have an abnormal rhythmof the heart. Patients must also not currently be taking osteoporosismedication. In addition, patients must not have hypocalcaemia orpres-existing dental diseases. All subjects are evaluated for safety andall blood collections for pharmacokinetic analysis are collected asscheduled. All studies are performed with institutional ethics committeeapproval and patient consent.

Phase I: Patients receive (e.g., intravenous, oral, ip, or the like)TNAP inhibitor (e.g., a compound of Formula I-IV, or a pharmaceuticallyacceptable salt thereof) daily for 4 weeks. Cohorts of 3-6 patientsreceive escalating doses of TNAP inhibitor (e.g., a compound of FormulaI-IV, or a pharmaceutically acceptable salt thereof). Escalation willnot be performed until all patients in the previous dose cohort havebeen treated for 4 weeks and until results obtained 4 weeks aftertreatment initiation do not reveal toxicity. Doses of TNAP inhibitor(e.g., a compound of Formula I-IV, or a pharmaceutically acceptable saltthereof) may be held or modified for toxicity based on assessments asoutlined below. Dose escalation is considered complete, if 2 patientsexperience a Grade 3 Adverse Event (AE) or if 1 patient experiencesGrade 4 AE at a particular cohort.

Phase II: Patients receive TNAP inhibitor (e.g., a compound of FormulaI-IV, or a pharmaceutically acceptable salt thereof) as in phase I at asuitable dose below the dose used in the final cohort. Treatmentcontinues throughout a 24-month study period during which clinical(which includes safety and tolerability) assessments are performed.

Blood Sampling: Serial blood is drawn by direct vein puncture before andafter administration TNAP inhibitor (e.g., a compound of Formula I-IV,or a pharmaceutically acceptable salt thereof). Venous blood samples (5mL) for determination of serum concentrations are obtained in-hospitalduring a 24-hour period. Each serum sample is divided into two aliquots.All serum samples are stored at −20° C. Serum samples are shipped on dryice.

Pharmacokinetics: Patients undergo plasma/serum sample collection forpharmacokinetic evaluation in-hospital during a 24-hour period.Pharmacokinetic parameters are calculated by model independent methodson a Digital Equipment Corporation VAX 8600 computer system using thelatest version of the BIOAVL software. The following pharmacokineticsparameters are determined: peak serum concentration (C_(max)); time topeak serum concentration (t_(max)); area under the concentration-timecurve (AUC) from time zero to the last blood sampling time (AUC₀₋₇₂)calculated with the use of the linear trapezoidal rule; and terminalelimination half-life (t_(1/2)), computed from the elimination rateconstant. The elimination rate constant is estimated by linearregression of consecutive data points in the terminal linear region ofthe log-linear concentration-time plot. The mean, standard deviation(SD), and coefficient of variation (CV) of the pharmacokineticparameters are calculated for each treatment. The ratio of the parametermeans (preserved formulation/non-preserved formulation) is calculated.

Patient Response: The primary outcome measure is safety andtolerability, based on conventional laboratory and clinical assessments.The secondary outcome measure is a change in elastic fibercalcification. A blinded dermatopathologist will grade skin biopsies onthe density of von Kossa staining. Other secondary outcome measuresinclude changes in skin lesions, and changes in disease progression,based on ophthalmologic examinations.

Example 10 Calciphylaxis Clinical Trial

Human Clinical Trial of the Safety and/or Efficacy of a TNAP inhibitor(e.g., a compound of Formula I-IV, or a pharmaceutically acceptable saltthereof) therapy.

Objective: To determine the safety, pharmacokinetics, and efficacy ofadministered TNAP inhibitor (e.g., a compound of Formula I-IV, or apharmaceutically acceptable salt thereof).

Study Design: This will be a Phase I, single-center, open-label,non-randomized dose escalation study followed by a Phase II study incalciphylaxis patients. The diagnosis of pseudoxanthoma elasticum mustbe confirmed by a skin biopsy or initial dermatology visit within theprevious 5 years, and a serum phosphorus level greater than 4.5 mg/dL.Patients must not have received other investigational agents within 3months of study initiation. Fertile patients must agree to use adequatecontraception throughout the study and for 18 months after cessation oftreatment with a TNAP inhibitor (e.g., a compound of Formula I-IV).Patients must not be undertaking renal replacement therapy. Patientsmust also not have had a recent fracture (within the last 3 months). Inaddition, patients must not have an abnormal rhythm of the heart.Patients must also not currently be taking osteoporosis medication. Inaddition, patients must not have hypocalcaemia or pres-existing dentaldiseases. All subjects are evaluated for safety and all bloodcollections for pharmacokinetic analysis are collected as scheduled. Allstudies are performed with institutional ethics committee approval andpatient consent.

Phase I: Patients receive (e.g., intravenous, oral, ip, or the like)TNAP inhibitor (e.g., a compound of Formula I-IV, or a pharmaceuticallyacceptable salt thereof) daily for 4 weeks. Cohorts of 3-6 patientsreceive escalating doses of TNAP inhibitor (e.g., a compound of FormulaI-IV, or a pharmaceutically acceptable salt thereof). Escalation willnot be performed until all patients in the previous dose cohort havebeen treated for 4 weeks and until results obtained 4 weeks aftertreatment initiation do not reveal toxicity. Doses of TNAP inhibitor(e.g., a compound of Formula I-IV, or a pharmaceutically acceptable saltthereof) may be held or modified for toxicity based on assessments asoutlined below. Dose escalation is considered complete, if 2 patientsexperience a Grade 3 Adverse Event (AE) or if 1 patient experiencesGrade 4 AE at a particular cohort.

Phase II: Patients receive TNAP inhibitor (e.g., a compound of FormulaI-IV, or a pharmaceutically acceptable salt thereof) as in phase I at asuitable dose below the dose used in the final cohort. Treatmentcontinues throughout a 24-month study period during which clinical(which includes safety and tolerability) assessments are performed.

Blood Sampling: Serial blood is drawn by direct vein puncture before andafter administration TNAP inhibitor (e.g., a a compound of Formula I-IV,or a pharmaceutically acceptable salt thereof). Venous blood samples (5mL) for determination of serum concentrations are obtained in-hospitalduring a 24-hour period. Each serum sample is divided into two aliquots.All serum samples are stored at −20° C. Serum samples are shipped on dryice.

Pharmacokinetics: Patients undergo plasma/serum sample collection forpharmacokinetic evaluation in-hospital during a 24-hour period.Pharmacokinetic parameters are calculated by model independent methodson a Digital Equipment Corporation VAX 8600 computer system using thelatest version of the BIOAVL software. The following pharmacokineticsparameters are determined: peak serum concentration (C_(max)); time topeak serum concentration (t_(max)); area under the concentration-timecurve (AUC) from time zero to the last blood sampling time (AUC₀₋₇₂)calculated with the use of the linear trapezoidal rule; and terminalelimination half-life (t_(1/2)), computed from the elimination rateconstant. The elimination rate constant is estimated by linearregression of consecutive data points in the terminal linear region ofthe log-linear concentration-time plot. The mean, standard deviation(SD), and coefficient of variation (CV) of the pharmacokineticparameters are calculated for each treatment. The ratio of the parametermeans (preserved formulation/non-preserved formulation) is calculated.

Patient Response: The primary outcome measure is safety andtolerability, based on conventional laboratory and clinical assessments.The secondary outcome measure is a change in elastic fibercalcification. A blinded dermatopathologist will grade skin biopsies onthe density of von Kossa staining. Other secondary outcome measuresinclude changes in skin lesions, and changes in disease progression,based on ophthalmologic examinations.

Example 11 Parenteral Composition of a Compound of Formula I-IV

To prepare a parenteral pharmaceutical composition suitable foradministration by injection, 100 mg of a compound of Formula I-IV, or awater soluble pharmaceutically acceptable salt thereof, is dissolved inDMSO and then mixed with 10 ml of 0.9% sterile saline solution. Themixture is incorporated into a dosage unit suitable for administrationby injection.

Example 12 Oral Composition of a Compound of Formula I-IV

To prepare a pharmaceutical composition for oral delivery, 400 mg of acompound of Formula I-IV and the following ingredients are mixedintimately and pressed into single scored tablets.

Tablet Formulation Quantity per tablet Ingredient mg compound 400cornstarch 50 croscarmellose sodium 25 lactose 120 magnesium stearate 5

The following ingredients are mixed intimately and loaded into ahard-shell gelatin capsule.

Capsule Formulation Quantity per capsule Ingredient mg compound 200lactose spray dried 148 magnesium stearate 2

While preferred embodiments of the present invention have been shown anddescribed herein, it will be obvious to those skilled in the art thatsuch embodiments are provided by way of example only. Numerousvariations, changes, and substitutions will now occur to those skilledin the art without departing from the invention. It should be understoodthat various alternatives to the embodiments of the invention describedherein may be employed in practicing the invention. It is intended thatthe following claims define the scope of the invention and that methodsand structures within the scope of these claims and their equivalents becovered thereby.

What is claimed is:
 1. A method of inhibiting tissue-nonspecificalkaline phosphatase (TNAP) activity in a subject with vascularcalcification, ectopic ossification in spinal ligaments, ankylosis,osteoarthritis, or pathological calcification comprising administering acompound of Formula I, or a pharmaceutically acceptable salt, polymorph,solvate, tautomer, or N-oxide thereof, to the subject in need thereof:

wherein: Y¹ is a bond, and Y² is —N(R⁶)—; L¹ and L² are each a bond; X¹is ═N— or ═C(R²)—; X² is ═N— or ═C(R³)—; R¹ and R⁴ are independentlyselected from the group consisting of —F, —Cl, —Br, —CN, —C(O)—N(R⁷)—R⁸,—C(O)—O—R⁹, —OMe, —OCF₃, optionally substituted phenyl, and optionallysubstituted 5- or 6-membered heteroaryl; R², R³, and R⁵ are hydrogen; R⁶is hydrogen; R⁷ and R⁸ are independently hydrogen, optionallysubstituted alkyl, haloalkyl, optionally substituted cycloalkyl,optionally substituted heterocycloalkyl, or optionally substitutedphenyl, or R⁷ and R⁸ together with the nitrogen atom to which they areattached form an optionally substituted heterocycloamino; R⁹ is selectedfrom the group consisting of hydrogen, optionally substituted alkyl,haloalkyl, optionally substituted cycloalkyl, optionally substitutedheterocycloalkyl, and optionally substituted phenyl; and A is—C(O)—N(R⁷)—R⁸, or —C(O)—O—R⁹; or A is an optionally substituted phenylor optionally substituted 5- or 6-membered heteroaryl, wherein A isselected from:

R¹² and R¹³ are independently selected from the group consisting ofhydrogen, halogen, —CN, —OH, —C(O)—O—R¹⁹, optionally substituted alkyl,optionally substituted cycloalkyl, optionally substitutedheterocycloalkyl, optionally substituted alkoxy, haloalkyl, haloalkoxy,optionally substituted phenyl, and optionally substituted 5- or6-membered heteroaryl; R¹⁹ is selected from the group consisting ofhydrogen, optionally substituted alkyl, haloalkyl, optionallysubstituted cycloalkyl, optionally substituted heterocycloalkyl, andoptionally substituted phenyl; and R¹⁵ is hydrogen or optionallysubstituted alkyl.
 2. The method of claim 1, wherein: the vascularcalcification is an arterial calcification.
 3. The method of claim 1,wherein: the vascular calcification is associated with diabetes mellitusI, diabetes mellitus II, idiopathic infantile arterial calcification(IIAC), Kawasaki disease, obesity, or increased age.
 4. The method ofclaim 1, wherein: the vascular calcification is associated with chronicrenal disease.
 5. The method of claim 1, wherein: the pathologicalcalcification is ankylosing spondylitis, tumoral calcinosis,fibrodysplasia ossificans progressiva, progressive osseous heteroplasia,pseudoxanthoma elasticum, ankylosis, osteoarthritis, general arterialcalcification in infancy (GACI), arterial calcification due todeficiency of CD73 (ACDC), Keutel syndrome, peritoneal calcification,heterotopic calcification in amputees, tibial artery calcification, bonemetastasis, prosthetic calcification, or Paget's disease of bone.
 6. Themethod of claim 1, wherein: Y¹ is a bond and Y² is —N(R⁶)—; X² is═C(R³)—; L¹ is a bond; L² is a bond; and R⁶ is hydrogen as shown inFormula Ie:


7. The method of claim 1, wherein: X¹ is ═C(R²)—.
 8. The method of claim1, wherein: R⁴ is optionally substituted phenyl, optionally substituted5- or 6-membered heteroaryl, —C(O)—N(R⁷)—R⁸, or —C(O)—O—R⁹.
 9. Themethod of claim 1, wherein: R¹ and R⁴ are independently selected fromthe group consisting of —F, —Cl, —Br, —CN, —OMe, and —OCF₃.
 10. Themethod of claim 1, wherein: A is optionally substituted phenyl oroptionally substituted 5- or 6-membered heteroaryl, wherein A isselected from:

R¹² and R¹³ are independently selected from the group consisting ofhydrogen, halogen, —CN, —OH, —C(O)—O—R¹⁹, optionally substituted alkyl,optionally substituted cycloalkyl, optionally substitutedheterocycloalkyl, optionally substituted alkoxy, haloalkyl, haloalkoxy,optionally substituted phenyl, and optionally substituted 5- or6-membered heteroaryl; R¹⁹ is selected from the group consisting ofhydrogen, optionally substituted alkyl, haloalkyl, optionallysubstituted cycloalkyl, optionally substituted heterocycloalkyl, andoptionally substituted phenyl; and R¹⁵ is hydrogen or optionallysubstituted alkyl.
 11. The method of claim 1, wherein: A is

 wherein R¹² and R¹³ are independently selected from the groupconsisting of hydrogen, —F, —CN, —OH, —OMe, and —C(O)—O—Me; or A is—C(O)—O—R⁹; or A is —C(O)—N(R⁷)—R⁸.
 12. The method of claim 11, wherein:A is —C(O)—O—R⁹, wherein R⁹ is selected from hydrogen, optionallysubstituted alkyl, optionally substituted cycloalkyl, and optionallysubstituted phenyl.
 13. The method of claim 11, wherein: A is—C(O)—N(R⁷)—R⁸, wherein R⁷ and R⁸ together with the nitrogen atom towhich they are attached form an optionally substituted heterocycloamino.14. The method of claim 13, wherein: the optionally substitutedheterocycloamino is an optionally substituted pyrrolidine, an optionallysubstituted piperidine, an optionally substituted morpholine, or anoptionally substituted piperazine.
 15. The method of claim 11, wherein:A is —C(O)—N(R⁷)—R⁸, wherein R⁷ is hydrogen and R⁸ is optionallysubstituted alkyl, optionally substituted cycloalkyl, or optionallysubstituted phenyl.
 16. The method of claim 11, wherein: A is—C(O)—N(R⁷)—R⁸, wherein R⁷ and R⁸ are hydrogen.
 17. A method ofinhibiting tissue-nonspecific alkaline phosphatase (TNAP) activity in asubject with vascular calcification, ectopic ossification in spinalligaments, ankylosis, osteoarthritis, or pathological calcificationcomprising administering one of the following compounds to the subjectin need thereof:

or a pharmaceutically acceptable salt thereof.